Pesticidal genes and methods of use

ABSTRACT

Compositions having pesticidal activity and methods for their use are provided. Compositions include isolated and recombinant polypeptides having pesticidal activity, recombinant and synthetic nucleic acid molecules encoding the polypeptides, DNA constructs and vectors comprising the nucleic acid molecules, host cells comprising the vectors, and antibodies to the polypeptides. Nucleotide sequences encoding the polypeptides can be used in DNA constructs or expression cassettes for transformation and expression in organisms of interest. The compositions and methods provided are useful for producing organisms with enhanced pest resistance or tolerance. Transgenic plants and seeds comprising a nucleotide sequence that encodes a pesticidal protein of the invention are also provided. Such plants are resistant to insects and other pests. Methods are provided for producing the various polypeptides disclosed herein, and for using those polypeptides for controlling or killing a pest. Methods and kits for detecting polypeptides of the invention in a sample are also included.

CROSS REFERENCE TO RELATED APPLICATION

This application is a divisional application of U.S. Nonprovisionalapplication Ser. No. 15/697,183, filed Sep. 6, 2017, which claims thebenefit of U.S. Provisional Application No. 62/383,773, filed Sep. 6,2016; U.S. Provisional Application No. 62/385,441, filed Sep. 9, 2016;U.S. Provisional Application No. 62/448,410, filed Jan. 20, 2017 andU.S. Provisional Application No. 62/477,036, filed Mar. 27, 2017, all ofwhich are hereby incorporated in their entireties by this reference.

FIELD

The invention is drawn to methods and compositions for controllingpests, particularly plant pests.

REFERENCE TO A SEQUENCE LISTING SUBMITTED AS A TEXT FILE VIA EFS-WEB

The official copy of the sequence listing is submitted electronicallyvia EFS-Web as an ASCII formatted sequence listing with a file namedAGB038US-SeqListing-1059489.txt, created on Sep. 6, 2017, and having asize of 1,001,165 bytes and is filed concurrently with thespecification. The sequence listing contained in this ASCII formatteddocument is part of the specification and is herein incorporated byreference in its entirety.

BACKGROUND

Pests, plant diseases, and weeds can be serious threats to crops. Lossesdue to pests and diseases have been estimated at 37% of the agriculturalproduction worldwide, with 13% due to insects, bacteria and otherorganisms.

Toxins are virulence determinants that play an important role inmicrobial pathogenicity and/or evasion of the host immune response.Toxins from the gram-positive bacterium Bacillus, particularly Bacillusthuringensis, have been used as insecticidal proteins. Currentstrategies use the genes expressing these toxins to produce transgeniccrops. Transgenic crops expressing insecticidal protein toxins are usedto combat crop damage from insects.

While the use of Bacillus toxins has been successful in controllinginsects, resistance to Bt toxins has developed in some target pests inmany parts of the world where such toxins have been used intensively.One way of solving this problem is sowing Bt crops with alternating rowsof regular non Bt crops (refuge). An alternative method to avoid or slowdown development of insect resistance is stacking insecticidal geneswith different modes of action against insects in transgenic plants. Thecurrent strategy of using transgenic crops expressing insecticidalprotein toxins is placing increasing emphasis on the discovery of noveltoxins, beyond those already derived from the bacterium Bacillusthuringiensis. These toxins may prove useful as alternatives to thosederived from B. thuringiensis for deployment in insect- andpest-resistant transgenic plants. Thus, new toxin proteins are needed.

SUMMARY

Compositions having pesticidal activity and methods for their use areprovided. Compositions include isolated and recombinant polypeptidesequences having pesticidal activity, recombinant and synthetic nucleicacid molecules encoding the pesticidal polypeptides, DNA constructscomprising the nucleic acid molecules, vectors comprising the nucleicacid molecules, host cells comprising the vectors, and antibodies to thepesticidal polypeptides. Nucleotide sequences encoding the polypeptidesprovided herein can be used in DNA constructs or expression cassettesfor transformation and expression in organisms of interest, includingmicroorganisms and plants.

The compositions and methods provided herein are useful for theproduction of organisms with enhanced pest resistance or tolerance.These organisms and compositions comprising the organisms are desirablefor agricultural purposes. Transgenic plants and seeds comprising anucleotide sequence that encodes a pesticidal protein of the inventionare also provided. Such plants are resistant to insects and other pests.

Methods are provided for producing the various polypeptides disclosedherein, and for using those polypeptides for controlling or killing apest. Methods and kits for detecting polypeptides of the invention in asample are also included.

DETAILED DESCRIPTION

The present inventions now will be described more fully hereinafter.These inventions may be embodied in many different forms and should notbe construed as limited to the embodiments set forth herein; rather,these embodiments are provided so that this disclosure will satisfyapplicable legal requirements. Like numbers refer to like elementsthroughout.

Many modifications and other embodiments of the inventions set forthherein will come to mind to one skilled in the art to which theseinventions pertain having the benefit of the teachings presented in theforegoing descriptions. Therefore, it is to be understood that theinventions are not to be limited to the specific embodiments disclosedand that modifications and other embodiments are intended to be includedwithin the scope of the appended claims. Although specific terms areemployed herein, they are used in a generic and descriptive sense onlyand not for purposes of limitation.

I. Polynucleotides and Polypeptides

Compositions and method for conferring pesticidal activity to anorganism are provided. The modified organism exhibits pesticidalresistance or tolerance. Recombinant pesticidal proteins, orpolypeptides and fragments and variants thereof that retain pesticidalactivity, are provided and include those set forth in SEQ ID NOs: 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58,59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76,77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109,110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123,124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137,138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151,152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165,166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179,180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193,194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207,208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220,221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234,235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248,249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262,263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276,277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290,291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304,305, 306, 307, 308, and/or 309. The pesticidal proteins are biologicallyactive (e.g., pesticidal) against pests including insects, fungi,nematodes, and the like. Nucleotides encoding the pesticidalpolypeptides, including for example, SEQ ID NOS: 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98,99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223,224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279,280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293,294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307,308, and/or 309 or active fragments or variants thereof, can be used toproduce transgenic organisms, such as plants and microorganisms. Thepesticidal proteins are biologically active (for example, arepesticidal) against pests including insects, fungi, nematodes, and thelike. Polynucleotides encoding the pesticidal polypeptides, includingfor example, SEQ ID NOS: 1-309 or active fragments or variants thereof,can be used to produce transgenic organisms, such as plants andmicroorganisms. The transformed organisms are characterized by genomesthat comprise at least one stably incorporated DNA construct comprisinga coding sequence for a pesticidal protein disclosed herein. In someembodiments, the coding sequence is operably linked to a promoter thatdrives expression of the encoded pesticidal polypeptide. Accordingly,transformed microorganisms, plant cells, plant tissues, plants, seeds,and plant parts are provided. A summary of various polypeptides, activevariants and fragments thereof, and polynucleotides encoding the sameare set forth below in Table 1. As noted in Table 1, various forms ofpolypeptides are provided. Full length pesticidal polypeptides, as wellas, modified versions of the original full-length sequence (i.e.,variants) are provided. Table 1 further denotes “CryBP1” sequences. Suchsequences (SEQ ID NOS: 213 and 86) comprise accessory polypeptides thatcan be associated with some of the toxin genes. In such instances, theCryBP1 sequences can be used alone or in combination with any of thepesticidal polypeptides provided herein. Table 1 further providesSplit-Cry C-terminus polypeptides. Such sequence comprise the sequenceof a downstream protein that has homology to the C-terminal end of theCry class of toxin genes and are usually found after a Cry gene that isnot full-length and is missing the expected C-terminal region.

TABLE 1 Summary of SEQ ID NOs, Gene Class, and Variants thereofPolypeptides of the invention (and polynucleotides encoding the same)Split-Cry include those having the Gene Full-length variant CryBP1C-terminus Gene % sequence identity Name SEQ ID No. SEQ ID No.(s) SEQ IDNo. SEQ ID No. Class listed below APG00589 1 MTX 75, 80, 85, 90, 95, 96,97, 98, 99 APG00737 2 MTX 95, 96, 97, 98, 99 APG00738 3 PI-PLC 70, 75,80, 85, 90, 95, 96, 97, 98, 99 APG00749 4 5 MTX 80, 85, 90, 95, 96, 97,98, 99 APG00769 6 MTX 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, 95, 96, 97, 98, 99 APG00788 7 MTX 99 APG00790 8 Cyt 50, 55, 60, 65,70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG00808 9 MTX 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG00809 10MTX 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG00906 11 MTX 80, 85, 90, 95, 96, 97, 98, 99 APG00945 12 Cyt 80, 85,90, 95, 96, 97, 98, 99 APG00955 13 14 MTX 70, 75, 80, 85, 90, 95, 96,97, 98, 99 APG00965 15 Cry 95, 96, 97, 98, 99 APG00974 16 17 Cry 75, 80,85, 90, 95, 96, 97, 98, 99 APG00989 18 19 MTX 70, 75, 80, 85, 90, 95,96, 97, 98, 99 APG01022 20 MTX 55, 60, 65, 70, 75, 80, 85, 90, 95, 96,97, 98, 99 APG01068 21 22 MTX 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG01078 23 MTX 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, 95, 96, 97, 98, 99 APG01084 24 Cry 35, 40, 45, 50, 55, 60, 65, 70,75, 80, 85, 90, 95, 96, 97, 98, 99 APG01103 25 MTX 95, 96, 97, 98, 99APG01121 26 Cyt 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG01150 27 MTX 100 APG01199 28 29 Cry 55, 60, 65, 70, 75, 80, 85, 90,95, 96, 97, 98, 99 APG01242 30 31 MTX 30, 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG01246 32 33 Cyt 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG01257 34 Cyt 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG01269 35 MTX 95,96, 97, 98, 99 APG01273 36 MTX 20, 25, 30, 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG01301 37 Cyt 55, 60, 65, 70,75, 80, 85, 90, 95, 96, 97, 98, 99 APG01401 38 MTX 40, 45, 50, 55, 60,65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG01420 39 40 MTX 95, 96,97, 98, 99 APG01451 41 MTX 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG01463 42 MTX 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96,97, 98, 99 APG01507 43 44 MTX 85, 90, 95, 96, 97, 98, 99 APG01508 45 46MTX 85, 90, 95, 96, 97, 98, 99 APG01536 47 48, 49 MTX 90, 95, 96, 97,98, 99 APG01700 50 51 Cry 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96,97, 98, 99 APG01882 52 53 MTX 30, 35, 40, 45, 50, 55, 60, 65, 70, 75,80, 85, 90, 95, 96, 97, 98, 99 APG01944 54 55 Cry 30, 35, 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG01992 56 57 Cry80, 85, 90, 95, 96, 97, 98, 99 APG02038 58 MTX 65, 70, 75, 80, 85, 90,95, 96, 97, 98, 99 APG02067 59 60, 61 213 Cry 90, 95, 96, 97, 98, 99APG02224 62 63 Cry 90, 95, 96, 97, 98, 99 APG02225 64 MTX 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG02280 65MTX 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG02387 66 67, 68Cry 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG02400 69 70MTX 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG02518 71 72MTX 90, 95, 96, 97, 98, 99 APG02531 73 74 MTX 30, 35, 40, 45, 50, 55,60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG02552 75 76 Cry 45,50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG02555 77 78MTX 95, 96, 97, 98, 99 APG02557 79 80 MTX 30, 35, 40, 45, 50, 55, 60,65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG02633 81 MTX 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG02643 8283 MTX 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90,95, 96, 97, 98, 99 APG02740 84 85 86 MTX 45, 50, 55, 60, 65, 70, 75, 80,85, 90, 95, 96, 97, 98, 99 APG02921 87 88 MTX 90, 95, 96, 97, 98, 99APG02923 89 Cyt 98, 99 APG03079 90 MTX 20, 25, 30, 35, 40, 45, 50, 55,60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG03114 91 92 MTX 85,90, 95, 96, 97, 98, 99 APG03238 93 94 Bin 95, 96, 97, 98, 99 APG03440 95Cry 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG03484 96 97, 98 MTX 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG03619 99 100 MTX 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90,95, 96, 97, 98, 99 APG03686 101 102 MTX 35, 40, 45, 50, 55, 60, 65, 70,75, 80, 85, 90, 95, 96, 97, 98, 99 APG03715 103 104 MTX 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG03747 105106 MTX 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98,99 APG03760 107 MTX 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG03831 108109 Bin 96, 97, 98, 99 APG03867 110 111 MTX 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG04067 112 113 MTX 35, 40, 45,50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG04099 114 115MTX 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG04152116 117 Cry 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96,97, 98, 99 APG04450 118 MTX 85, 90, 95, 96, 97, 98, 99 APG04483 119 MTX30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG04686 120 121 Bin 95, 96, 97, 98, 99 APG04721 122 Cry 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG04778 123MTX 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97,98, 99 APG04793 124 125 MTX 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, 95, 96, 97, 98, 99 APG04925 126 Cry 30, 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG05213 127 Bin 98, 99 APG05372128 129 MTX 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98,99 APG05500 130 131 MTX 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96,97, 98, 99 APG05553 132 Cry 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG05634 133 MTX 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG05706 134135 Cry 95, 96, 97, 98, 99 APG06001 136 137 MTX 25, 30, 35, 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG06281 138 MTX 15,20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97,98, 99 APG06324 139 140 MTX 90, 95, 96, 97, 98, 99 APG06338 141 142 Bin95, 96, 97, 98, 99 APG06381 143 144, 145 MTX 25, 30, 35, 40, 45, 50, 55,60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG06465 146 MTX 30, 35,40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG06501147 148 MTX 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96,97, 98, 99 APG06589 149 150 MTX 80, 85, 90, 95, 96, 97, 98, 99 APG06676151 MTX 75, 80, 85, 90, 95, 96, 97, 98, 99 APG06894 152 153 Cyt 90, 95,96, 97, 98, 99 APG06989 154 155, 156 MTX 90, 95, 96, 97, 98, 99 APG06997157 Cry 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG07002 158 159 MTX 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90,95, 96, 97, 98, 99 APG07020 160 161 Cry 55, 60, 65, 70, 75, 80, 85, 90,95, 96, 97, 98, 99 APG07114 162 163 MTX 30, 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG07220 164 Cry 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG07224 165166 Cry 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG07444 167 MTX 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95,96, 97, 98, 99 APG07445 168 169 Cry 20, 25, 30, 35, 40, 45, 50, 55, 60,65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG07470 170 Cry 15, 20, 25,30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG07639 171 MTX 95, 96, 97, 98, 99 APG07676 172 173 MTX 35, 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG07682 174 175 MTX35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG07738 176 177 MTX 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, 95, 96, 97, 98, 99 APG07780 178 MTX 25, 30, 35, 40, 45, 50, 55, 60,65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG07954 179 Cry 80, 85, 90,95, 96, 97, 98, 99 APG08029 180 181 MTX 30, 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG08138 182 MTX 30, 35, 40, 45,50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG08151 183 184Bin 95, 96, 97, 98, 99 APG08509 185 186 MTX 75, 80, 85, 90, 95, 96, 97,98, 99 APG08607 187 188, 189 Cry 35, 40, 45, 50, 55, 60, 65, 70, 75, 80,85, 90, 95, 96, 97, 98, 99 APG08628 190 191 Cry 25, 30, 35, 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG08780 192 193 Cyt95, 96, 97, 98, 99 APG08794 194 195 MTX 30, 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG09055 196 197 MTX 65, 70, 75,80, 85, 90, 95, 96, 97, 98, 99 APG09096 198 199 Cry22 45, 50, 55, 60,65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG09376 200 201 Cry 95, 96,97, 98, 99 APG09455 202 203 MTX 25, 30, 35, 40, 45, 50, 55, 60, 65, 70,75, 80, 85, 90, 95, 96, 97, 98, 99 APG09642 204 205 MTX 35, 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG09659 206 207 Cry15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96,97, 98, 99 APG09717 208 Cyt 95, 96, 97, 98, 99 APG09726 209 210 MTX 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG09735 211 212 MTX35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG00743 214 215 MTX 97, 98, 99 APG01705 216 217 MTX 85, 90, 95, 96, 97,98, 99 APG01989 218 Cyt 100 APG02245 219 MTX 97, 98, 99 APG02279 220 221MTX 95, 96, 97, 98, 99 APG02429 222 MTX 97, 98, 99 APG02674 223 224 MTX97, 98, 99 APG02768 225 MTX 97, 98, 99 APG03040 226 MTX 99 APG03185 227228 MTX 90, 95, 96, 97, 98, 99 APG03217 229 230 MTX 30, 35, 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG03368 231 232 MTX90, 95, 96, 97, 98, 99 APG03662 233 234 MTX 65, 70, 75, 80, 85, 90, 95,96, 97, 98, 99 APG04224 235 236 MTX 80, 85, 90, 95, 96, 97, 98, 99APG04226 237 238 MTX 90, 95, 96, 97, 98, 99 APG04485 239 240 MTX 90, 95,96, 97, 98, 99 APG04643 241 242, 243 MTX 95, 96, 97, 98, 99 APG05660 244245 Cry 97, 98, 99 APG05969 246 MTX 90, 95, 96, 97, 98, 99 APG06528 247248 249 Cry 95, 96, 97, 98, 99 APG07049 250 251 MTX 90, 95, 96, 97, 98,99 APG07574 252 253 MTX 90, 95, 96, 97, 98, 99 APG08043 254 255 MTX 25,30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99APG08085 256 257 MTX 95, 96, 97, 98, 99 APG08225 258 259 MTX 98, 99APG08241 260 MTX 96, 97, 98, 99 APG08411 261 262 MTX 90, 95, 96, 97, 98,99 APG08718 263 MTX 100 APG08973 264 265 Cry 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95, 96, 97, 98, 99 APG08990 266 267 MTX 90, 95, 96,97, 98, 99 APG09256 268 MTX 96, 97, 98, 99 APG09842 269 MTX 98, 99APG02960 270 271, 272 MTX 95, 96, 97, 98, 99 APG01265.0 273 MTX 96, 97,98, 99, 100 APG03954.0 274 275 MTX 99, 100 APG04375.0 276 277 MTX 85,90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 APG04900.0 278 279 MTX 35,40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97,98, 99, 100 APG00770.0 280 Cyt 95, 96, 97, 98, 99, 100 APG01577.0 281MTX 95, 96, 97, 98, 99, 100 APG01797.0 282 MTX 5, 10, 15, 20, 25, 30,35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96,97, 98, 99, 100 APG01987.0 283 Cyt 98, 99, 100 APG02248.0 284 MTX 93,94, 95, 96, 97, 98, 99, 100 APG02443.0 285 MTX 90, 91, 92, 93, 94, 95,96, 97, 98, 99, 100 APG02756.0 286 MTX 95, 96, 97, 98, 99, 100APG03017.0 287 MTX 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100APG03148.0 288 MTX 94, 95, 96, 97, 98, 99, 100 APG03574.0 289 290 Bin45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98,99, 100 APG05399.0 291 MTX 91, 92, 93, 94, 95, 96, 97, 98, 99, 100APG05678.0 292 MTX 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100APG05707.0 293 Cyt 97, 98, 99, 100 APG06385.0 294 Cyt 91, 92, 93, 94,95, 96, 97, 98, 99, 100 APG08372.0 295 296 MTX 50, 55, 60, 65, 70, 75,80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 APG09444.0 297 298Cyt 95, 96, 97, 98, 99, 100 APG09446.0 299 MTX 90, 91, 92, 93, 94, 95,96, 97, 98, 99, 100 APG09857.0 300 MTX 93, 94, 95, 96, 97, 98, 99, 100APG09892.0 301 MTX 93, 94, 95, 96, 97, 98, 99, 100 APG01172 302 303, 304305 Cry 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96,97, 98, 99 APG01288 306 309 Cry 30, 35, 40, 45, 50, 55, 60, 65, 70, 75,80, 85, 90, 95, 96, 97, 98, 99 APG05711 307 308 Cry 40, 45, 50, 55, 60,65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 Polypeptides of the invention(and polynucleotides encoding the same) include those having Gene thesimilarity set Name forth below Homologs APG00589 85, 90, 95, 96, 97,APG00137 98, 99 (86.56% identity, 91.15% similarity) APG00345 (73.58%identity, 84.95% similarity) J8YPM2_BACCE (71.48% identity, 83.22%similarity) APG07639 (71.48% identity, 81.88% similarity) CA_2844913-100(70.81% identity, 82.55% similarity) APG00107 (69.80% identity, 83.56%similarity) APG00201 (67.53% identity, 79.22% similarity) APG00847(67.53% identity, 78.90% similarity) APG00749 (67.44% identity, 80.07%similarity) APG00955 (64.14% identity, 75.00% similarity) APG01451(57.05% identity, 66.78% similarity) APG00737 97, 98, 99 APG01269(96.06% identity, 97.24% similarity) WP_044585299.1 (94.86% identity,96.84% similarity) WP_000586615.1 (94.47% identity, 96.05% similarity)WP_000586617.1 (94.07% identity, 96.44% similarity) APG01103 (93.68%identity, 97.23% similarity) APG01150 (83.40% identity, 90.51%similarity) APG00788 (82.21% identity, 89.72% similarity) Cry46Ab(29.64% identity, 44.95% similarity) APG00738 80, 85, 90, 95, 96,APG00638 97, 98, 99 (68.73% identity, 80.08% similarity) WP_050845433.1(66.21% identity, 77.27% similarity) WP_000513490.1 (61.75% identity,70.87% similarity) AGA40046.1 (60.97% identity, 72.04% similarity)APG00749 90, 95, 96, 97, 98, APG00155 99 (81.79% identity, 88.08%similarity) APG00006 (79.34% identity, 89.51% similarity) APG00201(79.02% identity, 87.54% similarity) APG00566 (78.76% identity, 88.89%similarity) APG00847 (78.69% identity, 88.20% similarity) APG07639(77.00% identity, 84.33% similarity) J8YPM2_BACCE (76.74% identity,85.38% similarity) CA_2844913-100 (75.75% identity, 84.72% similarity)APG00955 (70.16% identity, 81.64% similarity) APG01451 (68.00% identity,72.00% similarity) APG00589 (67.44% identity, 80.07% similarity)APG00769 45, 50, 55, 60, 65, APG00535 70, 75, 80, 85, 90, (73.91%identity, 95, 96, 97, 98, 99 83.28% similarity) APG00016 (73.02%identity, 80.00% similarity) ANN35810.1 (25.87% identity, 42.27%similarity) Cry23Aa1 (25.79% identity, 40.25% similarity) APG00788 99APG01150 (98.42% identity, 98.81% similarity) WP_061530406.1 (98.42%identity, 98.81% similarity) WP_064474064.1 (97.63% identity, 98.02%similarity) SCA96918.1 (96.44% identity, 97.23% similarity) APG00737(82.21% identity, 89.72% similarity) APG01103 (81.42% identity, 89.33%similarity) APG01269 (80.31% identity, 88.58% similarity) APG00790 70,75, 80, 85, 90, APG01257 95, 96, 97, 98, 99 (92.91% identity, 95.39%similarity) APG00431 (81.56% identity, 91.13% similarity) APG00015(61.27% identity, 76.76% similarity) APG00468 (60.92% identity, 76.06%similarity) APG01121 (59.86% identity, 75.00% similarity) APG01301(59.39% identity, 75.77% similarity) APG00168 (51.19% identity, 70.17%similarity) CT2BB_BACTJ (45.73% identity, 65.19% similarity) CAC80987.1(44.64% identity, 67.47% similarity) Cyt2Ba10 (42.81% identity, 65.41%similarity) APG00808 40, 45, 50, 55, 60, AGA40031.1 65, 70, 75, 80, 85,(21.54% identity, 90, 95, 96, 97, 98, 37.85% similarity) 99 CAA63374.1(21.24% identity, 38.05% similarity) AGA40029.1 (20.72% identity, 36.94%similarity) US_8829279_B2-35 (20.36% identity, 36.53% similarity)APG00809 55, 60, 65, 70, 75, APG01401 80, 85, 90, 95, 96, (99.67%identity, 97, 98, 99 100.00% similarity) US_2016_0031949_A1-30 (36.88%identity, 53.49% similarity) US_2016_0031949_A1-36 (36.88% identity,53.49% similarity) WP_024361905.1 (36.00% identity, 53.67% similarity)Cry46Aa2 (34.01% identity, 50.43% similarity) APG00906 90, 95, 96, 97,98, WP_016096697.1 99 (77.17% identity, 87.40% similarity) SCA96918.1(76.77% identity, 87.80% similarity) WP_064474064.1 (76.38% identity,87.80% similarity) APG01150 (76.38% identity, 87.40% similarity)APG01269 (71.65% identity, 85.83% similarity) APG00737 (71.26% identity,85.04% similarity) APG01103 (71.26% identity, 85.04% similarity)APG00945 90, 95, 96, 97, 98, WP_046912431.1 99 (76.88% identity, 82.41%similarity) WP_019890121.1 (76.65% identity, 85.28% similarity)WP_030212110.1 (75.13% identity, 86.80% similarity) APG08780 (74.87%identity, 81.91% similarity) APG00134 (74.62% identity, 84.77%similarity) APG06894 (70.41% identity, 84.69% similarity) APG00327(58.85% identity, 64.62% similarity) APG00108 (53.87% identity, 61.62%similarity) APG00955 80, 85, 90, 95, 96, APG00749 97, 98, 99 (70.16%identity, 81.64% similarity) APG00201 (68.28% identity, 78.96%similarity) APG00847 (66.88% identity, 78.25% similarity) APG00260(66.45% identity, 78.39% similarity) APG00006 (66.24% identity, 77.39%similarity) CA_2844913-100 (65.58% identity, 75.97% similarity)J8YPM2_BACCE (65.58% identity, 75.32% similarity) APG07639 (65.48%identity, 74.52% similarity) APG00566 (65.40% identity, 78.10%similarity) APG00589 (64.14% identity, 75.00% similarity) APG01451(57.57% identity, 66.45% similarity) APG00965 96, 97, 98, 99WP_001087511.1 (91.04% identity, 95.15% similarity) CA_2844913-12(91.04% identity, 94.86% similarity) APG07954 (86.42% identity, 90.22%similarity) EEM92570.1 (84.14% identity, 88.25% similarity) APG00152(70.43% identity, 81.45% similarity) APG00084 (57.12% identity, 68.83%similarity) APG00974 80, 85, 90, 95, 96, APG00068 - 97, 98, 99US_2016_0177333_A1-35 (78.02% identity, 86.40% similarity) APG00310(74.58% identity, 83.11% similarity) US_2016_0017363_A1-32 (70.44%identity, 78.98% similarity) APG00054 (70.13% identity, 78.91%similarity) US_2016_0017363_A1-33 (69.9% identity, 78.21% similarity)APG00673 (60.65% identity, 70.20% similarity) Cry32Aa2 (56.80% identity,65.59% similarity) APG00105 (54.82% identity, 65.27% similarity)APG00989 85, 90, 95, 96, 97, APG00309 98, 99 (79.88% identity, 87.69%similarity) C3HSG6_BACTU (67.27% identity, 80.00% similarity)WP_018673409.1 (63.17% identity, 76.65% similarity) APG00585 (61.68%identity, 75.45% similarity) APG00427 (61.52% identity, 75.45%similarity) APG00851 (60.30% identity, 74.24% similarity) WP_061663532.1(60.30% identity, 74.24% similarity) US_2008_0070829_A1-2 (51.36%identity, 67.98% similarity) APG01022 65, 70, 75, 80, 85, APG02400 90,95, 96, 97, 98, (99.73% identity, 99 100.00% similarity) APG00426(97.32% identity, 98.12% similarity) APG00891 (92.56% identity, 93.33%similarity) APG00227 (90.75% identity, 91.00% similarity) APG00517(81.23% identity, 82.84% similarity) APG00158 (64.72% identity, 72.68%similarity) APG01508 (57.18% identity, 69.95% similarity) J8H0D9_BACCE(52.51% identity, 62.06% similarity) WP_063226258.1 (46.97% identity,60.16% similarity) APG01068 80, 85, 90, 95, 96, WP_003210059.1 97, 98,99 (64.53% identity, 77.62% similarity) WP_002107243.1 (58.62% identity,73.56% similarity) WP_003209347.1 (36.83% identity, 51.84% similarity)WP_006097189.1 (36.57% identity, 52.86% similarity) APG01078 25, 30, 35,40, 45, AGA40044.1 50, 55, 60, 65, 70, (16.91% identity, 75, 80, 85, 90,95, 23.32% similarity) 96, 97, 98, 99 APG01084 50, 55, 60, 65, 70,APG00603 75, 80, 85, 90, 95, (87.45% identity, 96, 97, 98, 99 93.21%similarity) APG01112 (71.19% identity, 76.34% similarity) WP_017762616.1(32.52% identity, 49.48% similarity) WP_044306756.1 (30.71% identity,47.57% similarity) WP_048536362.1 (28.87% identity, 42.58% similarity)AGA40058.1 (27.63% identity, 40.59% similarity) APG01103 99WP_044585299.1 (94.86% identity, 98.81% similarity) WP_000586617.1(94.86% identity, 98.42% similarity) WP_061667036.1 (94.47% identity,98.81% similarity) APG00737 (93.68% identity, 97.23% similarity)APG01269 (93.31% identity, 97.64% similarity) APG01150 (82.61% identity,90.12% similarity) APG00788 (81.42% identity, 89.33% similarity)APG01121 70, 75, 80, 85, 90, APG01301 95, 96, 97, 98, 99 (86.12%identity, 93.24% similarity) APG00468 (80.07% identity, 89.30%similarity) APG00015 (80.07% identity, 88.93% similarity) APG00168(64.73% identity, 74.32% similarity) APG01257 (59.93% identity, 76.24%similarity) APG00790 (59.86% identity, 75.00% similarity) APG00431(58.80% identity, 75.35% similarity) ACF35049.1 (53.24% identity, 68.35%similarity) CA_2866166-1433 (53.24% identity, 68.35% similarity)AF398463_1 (51.66% identity, 64.58% similarity) Cyt2Bc1 (49.82%identity, 65.72% similarity) APG01150 100 KXY42528.1 (100.00% identity,100.00% similarity) APG00788 (98.42% identity, 98.81% similarity)WP_064474064.1 (98.42% identity, 98.42% similarity) SCA96918.1 (97.23%identity, 97.63% similarity) APG00737 (83.40% identity, 90.51%similarity) APG01103 (82.61% identity, 90.12% similarity) APG01269(81.50% identity, 89.37% similarity) APG01199 70, 75, 80, 85, 90,US_8759619_B2-26 95, 96, 97, 98, 99 (54.49% identity, 66.62% similarity)ADK91079.1 (47.90% identity, 61.76% similarity) AGP18059.1 (47.77%identity, 61.59% similarity) APG01242 45, 50, 55, 60, 65, AGP17979.1 70,75, 80, 85, 90, (28.88% identity, 95, 96, 97, 98, 99 44.39% similarity)WP_061688274.1 (27.99% identity, 42.49% similarity)US_2016_0017363_A1-60 (26.11% identity, 42.30% similarity)US_2016_0017363_A1-61 (25.59% identity, 41.51% similarity) APG01246 60,65, 70, 75, 80, CT2BB_BACTJ 85, 90, 95, 96, 97, (39.42% identity, 98, 9955.47% similarity) CA_2618430-14 (38.61% identity, 54.05% similarity)Cyt2Aa2 (38.29% identity, 54.65% similarity) APG01257 70, 75, 80, 85,90, APG00790 95, 96, 97, 98, 99 (92.91% identity, 95.39% similarity)APG00431 (82.98% identity, 91.13% similarity) APG00015 (61.35% identity,76.24% similarity) APG00468 (60.99% identity, 76.60% similarity)APG01121 (59.93% identity, 76.24% similarity) APG01301 (58.82% identity,75.09% similarity) APG00168 (50.67% identity, 68.79% similarity)ACF35049.1 (46.50% identity, 67.13% similarity) CA_2866166-1433 (46.50%identity, 67.13% similarity) US_2013_0097729_A1-40 (46.37% identity,66.78% similarity) Cyt2Ba15 (44.52% identity, 66.10% similarity)APG01269 97, 98, 99 APG00737 (96.06% identity, 97.24% similarity)WP_061667036.1 (94.49% identity, 96.85% similarity) ADQ73630.1 (94.49%identity, 96.46% similarity) WP_000586614.1 (94.09% identity, 96.46%similarity) APG01103 (93.31% identity, 97.64% similarity) APG01150(81.50% identity, 89.37% similarity) APG00788 (80.31% identity, 88.58%similarity) APG01273 35, 40, 45, 50, 55, APG07780 60, 65, 70, 75, 80,(70.07% identity, 85, 90, 95, 96, 97, 82.04% similarity) 98, 99WP_033694890.1 (19.47% identity, 32.63% similarity) ANN35812.1 (16.27%identity, 26.42% similarity) APG01301 70, 75, 80, 85, 90, APG01121 95,96, 97, 98, 99 (86.12% identity, 93.24% similarity) APG00468 (80.07%identity, 87.90% similarity) APG00015 (79.36% identity, 87.19%similarity) APG00168 (63.61% identity, 74.83% similarity) APG00431(59.79% identity, 76.63% similarity) APG00790 (59.39% identity, 75.77%similarity) APG01257 (58.82% identity, 75.09% similarity) ACF35049.1(54.77% identity, 68.90% similarity) Cyt2Aa2 (54.77% identity, 68.9%similarity) APG01401 55, 60, 65, 70, 75, APG00809 80, 85, 90, 95, 96,(99.67% identity, 97, 98, 99 100.00% similarity) US_2016_0031949_A1-30(37.21% identity, 53.49% similarity) WP_024361905.1 (36.33% identity,53.67% similarity) BAD35170.1 (36.28% identity, 51.83% similarity)Cry46Aa2 (34.29% identity, 50.43% similarity) APG01420 96, 97, 98, 99WP_061688274.1 (93.10% identity, 95.69% similarity) WP_050595402.1(89.94% identity, 91.67% similarity) WP_023524027.1 (50.42% identity,65.37% similarity) Cry55Aa1 (21.61% identity, 35.43% similarity)APG01451 75, 80, 85, 90, 95, APG00155 96, 97, 98, 99 (77.70% identity,79.39% similarity) APG00749 (68.00% identity, 72.00% similarity)APG00107 (67.23% identity, 74.66% similarity) APG07639 (67.00% identity,72.39% similarity) J8YPM2_BACCE (66.55% identity, 73.31% similarity)APG00930 (65.88% identity, 74.32% similarity) CA_2844913-100 (64.86%identity, 72.30% similarity) APG00201 (64.14% identity, 70.39%similarity) APG00847 (63.82% identity, 70.39% similarity) APG00955(57.57% identity, 66.45% similarity) APG00589 (57.05% identity, 66.78%similarity) APG01463 50, 55, 60, 65, 70, WP_004999216.1 75, 80, 85, 90,95, (33.82% identity, 96, 97, 98, 99 49.12% similarity) WP_027393890.1(33.53% identity, 49.41% similarity) WP_015268242.1 (31.87% identity,45.91% similarity) WP_038918640.1 (31.10% identity, 46.04% similarity)APG01507 95, 96, 97, 98, 99 APG04450 (85.58% identity, 92.63%similarity) APG06589 (84.84% identity, 87.17% similarity) WP_016131662.1(82.90% identity, 92.90% similarity) WP_016132978.1 (82.58% identity,92.90% similarity) WP_016113347.1 (82.58% identity, 92.58% similarity)APG02921 (81.41% identity, 89.74% similarity) APG06989 (80.45% identity,89.74% similarity) Cry55Aa1 (18.86% identity, 34.74% similarity)APG01508 85, 90, 95, 96, 97, APG00020 98, 99 (98.32% identity, 99.44%similarity) APG00253 (89.39% identity, 94.13% similarity) APG00418(86.59% identity, 90.78% similarity) APG00764 (83.78% identity, 88.03%similarity) APG00646 (82.45% identity, 87.77% similarity) J8H0D9_BACCE(80.59% identity, 83.51% similarity) APG01022 (57.18% identity, 69.95%similarity) APG02400 (57.18% identity, 69.95% similarity) WP_063226258.1(49.87% identity, 63.00% similarity) CA_2844913-110 (46.17% identity,63.66% similarity) APG01536 95, 96, 97, 98, 99 APG02921 (89.42%identity, 93.91% similarity) APG06989 (88.46% identity, 93.59%similarity) WP_025150761.1 (86.86% identity, 92.63% similarity) APG01507(83.39% identity, 90.10% similarity) WP_016131662.1 (80.83% identity,89.46% similarity) WP_016132978.1 (80.19% identity, 89.46% similarity)APG06589 (74.86% identity, 81.50% similarity) APG01700 70, 75, 80, 85,90, EWY82070.1 95, 96, 97, 98, 99 (49.25% identity, 65.86% similarity)EXM20686.1 (47.95% identity, 64.74% similarity) EXK36684.1 (47.95%identity, 64.55% similarity) EXL68750.1 (46.64% identity, 62.69%similarity) APG01882 45, 50, 55, 60, 65, APG00696 70, 75, 80, 85, 90,(97.06% identity, 95, 96, 97, 98, 99 97.06% similarity) APG08138 (96.73%identity, 97.71% similarity) APG06465 (95.75% identity, 97.39%similarity) APG04483 (95.75% identity, 97.06% similarity) APG03715(92.19% identity, 93.13% similarity) US_2015_0047076_A1-6 (25.55%identity, 44.48% similarity) AGP18056.1 (25.00% identity, 41.88%similarity) US_2012_0278954_A1-26 (24.77% identity, 43.20% similarity)Cry45Aa (23.93% identity, 41.41% similarity) APG01944 35, 40, 45, 50,55, WP_057506999.1 60, 65, 70, 75, 80, (26.27% identity, 85, 90, 95, 96,97, 33.13% similarity) 98, 99 APG01992 90, 95, 96, 97, 98, ABU96490.1 99(79.97% identity, 85.94% similarity) ACP43735.1 (72.82% identity, 79.06%similarity) AJW76683.1 (58.47% identity, 70.48% similarity) APG02038 75,80, 85, 90, 95, APG00661 96, 97, 98, 99 (60.90% identity, 73.43%similarity) US_8829279_B2-25 (60.78% identity, 72.75% similarity)AGA40029.1 (59.88% identity, 73.25% similarity) US_8829279_B2-35 (58.31%identity, 71.90% similarity) APG00528 (55.29% identity, 63.06%similarity) AGA40031.1 (51.24% identity, 67.7% similarity) APG02067 95,96, 97, 98, 99 R8F389_BACCE (87.78% identity, 90.66% similarity)R8DGL6_BACCE (87.70% identity, 90.66% similarity) APG00059 -US_2016_0177333_A1-27 (65.06% identity, 74.02% similarity) Cry69Aa1(61.62% identity, 72.66% similarity) APG00662 (57.80% identity, 70.02%similarity) APG00079 (54.27% identity, 67.02% similarity) APG00786(53.80% identity, 65.83% similarity) APG02224 95, 96, 97, 98, 99CA_2866166-476 (85.51% identity, 91.42% similarity) AAR98783.1 (85.42%identity, 91.42% similarity) ADB54826.1 (85.42% identity, 91.34%similarity) Cry8Na1 (70.61% identity, 81.02% similarity) APG02225 45,50, 55, 60, 65, AGA40058.1 70, 75, 80, 85, 90, (28.80% identity, 95, 96,97, 98, 99 41.87% similarity) US_2013_0227743_A1-200 (25.67% identity,35.83% similarity) WP_049523612.1 (24.85% identity, 38.97% similarity)WP_033684369.1 (23.79% identity, 35.50% similarity) APG02280 75, 80, 85,90, 95, APG00846 96, 97, 98, 99 (81.66% identity, 87.97% similarity)APG00513 (79.08% identity, 87.68% similarity) AGA40030.1 (57.46%identity, 72.10% similarity) APG05634 (54.83% identity, 70.17%similarity) APG03760 (53.30% identity, 71.06% similarity)US_8829279_B2-27 (52.99% identity, 65.24% similarity) APG00224 (52.97%identity, 69.69% similarity) APG00609 (51.39% identity, 67.50%similarity) APG02387 60, 65, 70, 75, 80, AGP18054.1 85, 90, 95, 96, 97,(50.77% identity, 98, 99 59.76% similarity) R8EX84_BACCE (37.32%identity, 51.11% similarity) APG02400 65, 70, 75, 80, 85, APG01022 90,95, 96, 97, 98, (99.73% identity, 99 100.00% similarity) APG00426(97.05% identity, 98.12% similarity) APG00891 (92.56% identity, 93.33%similarity) APG00227 (90.75% identity, 91.00% similarity) APG00517(81.23% identity, 82.84% similarity) APG00158 (64.72% identity, 72.68%similarity) APG01508 (57.18% identity, 69.95% similarity) J8H0D9_BACCE(52.51% identity, 62.06% similarity) WP_063226258.1 (46.97% identity,60.16% similarity) APG02518 95, 96, 97, 98, 99 WP_047426867.1 (88.48%identity, 93.64% similarity) WP_062673074.1 (86.71% identity, 93.05%similarity) WP_034734550.1 (77.95% identity, 90.33% similarity) APG03114(77.58% identity, 89.39% similarity) WP_027372020.1 (77.34% identity,89.73% similarity) APG02531 45, 50, 55, 60, 65, APG07114 70, 75, 80, 85,90, (94.10% identity, 95, 96, 97, 98, 99 95.28% similarity) APG04067(67.84% identity, 81.29% similarity) CA_2844913-114 (28.39% identity,44.01% similarity) AIK29697.1 (26.85% identity, 40.92% similarity)ADK08315.1 (25.80% identity, 39.89% similarity) KOS27986.1 (24.86%identity, 38.11% similarity) APG02552 60, 65, 70, 75, 80, WP_048179486.185, 90, 95, 96, 97, (41.38% identity, 98, 99 57.09% similarity)WP_026632766.1 (34.53% identity, 48.84% similarity) APG02555 99WP_044444098.1 (94.72% identity, 98.02% similarity) WP_060749709.1(88.82% identity, 94.41% similarity) APG01507 (55.56% identity, 72.06%similarity) WP_016132978.1 (53.94% identity, 69.40% similarity) APG02921(53.92% identity, 70.22% similarity) APG06989 (53.31% identity, 70.35%similarity) APG02557 45, 50, 55, 60, 65, APG00994 70, 75, 80, 85, 90,(57.22% identity, 95, 96, 97, 98, 99 72.75% similarity) APG00429 (56.32%identity, 72.25% similarity) US_8796026_B2-33 (26.47% identity, 43.14%similarity) US_8796026_B2-10 (26.00% identity, 43.50% similarity)US_8796026_B2-8 (24.82% identity, 41.12% similarity) WP_019419944.1(23.82% identity, 38.68% similarity) APG02633 45, 50, 55, 60, 65,US_2015_0047076_A1-12 70, 75, 80, 85, 90, (27.61% identity, 95, 96, 97,98, 99 42.76% similarity) WP_029440439.1 (25.43% identity, 41.92%similarity) US_2012_0278954_A1-30 (24.20% identity, 44.13% similarity)Cry23Aa1 (23.49% identity, 44.13% similarity) APG02643 20, 25, 30, 35,40, BAC78489.1 45, 50, 55, 60, 65, (14.20% identity, 70, 75, 80, 85, 90,19.14% similarity) 95, 96, 97, 98, 99 1W3AA (12.96% identity, 18.52%similarity) BAC78490.1 (12.96% identity, 18.21% similarity) EPT04861.1(12.54% identity, 16.81% similarity) APG02740 65, 70, 75, 80, 85,AAA22332.1 90, 95, 96, 97, 98, (43.34% identity, 99 62.54% similarity)AF316145_1 (31.25% identity, 54.51% similarity) ADY24997.2 (28.62%identity, 44.48% similarity) WP_033694890.1 (27.59% identity, 41.64%similarity) APG02921 95, 96, 97, 98, 99 APG06989 (97.76% identity,98.40% similarity) WP_025150761.1 (89.74% identity, 93.91% similarity)APG01536 (89.42% identity, 93.91% similarity) WP_016131662.1 (82.69%identity, 89.42% similarity) WP_016132978.1 (82.37% identity, 89.74%similarity) APG01507 (81.41% identity, 89.74% similarity) APG06589(73.91% identity, 80.87% similarity) APG02923 99 WP_016110459.1 (97.05%identity, 98.31% similarity) APG00128 (93.25% identity, 97.05%similarity) APG00177 (56.25% identity, 69.53% similarity) APG00126(54.40% identity, 69.60% similarity) APG00437 (53.46% identity, 68.08%similarity) APG00121 (52.52% identity, 62.59% similarity) WP_016110460.1(52.19% identity, 68.13% similarity) APG03079 25, 30, 35, 40, 45,AGP17988.1 50, 55, 60, 65, 70, (18.22% identity, 75, 80, 85, 90, 95,24.45% similarity) 96, 97, 98, 99 APG03114 95, 96, 97, 98, 99WP_034734550.1 (83.28% identity, 92.40% similarity) WP_027372020.1(82.98% identity, 91.79% similarity) WP_061085076.1 (81.76% identity,90.88% similarity) WP_047426867.1 (78.98% identity, 90.69% similarity)APG02518 (77.58% identity, 89.39% similarity) APG03238 95, 96, 97, 98,99 APG04686 (98.50% identity, 99.14% similarity) APG08151 (97.86%identity, 98.72% similarity) APG03831 (97.00% identity, 98.29%similarity) APG06338 (96.79% identity, 98.07% similarity) KIF65774.1(93.36% identity, 94.65% similarity) KHA73845.1 (70.02% identity, 79.87%similarity) WP_042557199.1 (59.28% identity, 69.94% similarity)WP_064117037.1 (56.81% identity, 68.09% similarity) APG03440 50, 55, 60,65, 70, WP_044306759.1 75, 80, 85, 90, 95, (30.62% identity, 96, 97, 98,99 44.66% similarity) AGP17992.1 (30.62% identity, 44.50% similarity)WP_017762581.1 (30.59% identity, 45.65% similarity) AGA40058.1 (28.89%identity, 42.74% similarity) APG03484 70, 75, 80, 85, 90, APG00366 95,96, 97, 98, 99 (79.41% identity, 86.20% similarity) W8YCZ9_BACTU (58.01%identity, 69.53% similarity) WP_061657790.1 (58.01% identity, 69.53%similarity) APG00129 (57.62% identity, 69.06% similarity) APG00681(52.12% identity, 66.59% similarity) APG03619 40, 45, 50, 55, 60,WP_065486138.1 65, 70, 75, 80, 85, (26.69% identity, 90, 95, 96, 97, 98,35.78% similarity) 99 WP_065487080.1 (26.69% identity, 35.78%similarity) AGS78124.1 (22.04% identity, 30.30% similarity) APG03686 50,55, 60, 65, 70, APG04793 75, 80, 85, 90, 95, (96.25% identity, 96, 97,98, 99 98.13% similarity) APG07676 (89.14% identity, 94.38% similarity)APG03747 (87.64% identity, 94.01% similarity) APG00014 (50.37% identity,66.67% similarity) WP_033694890.1 (34.63% identity, 46.54% similarity)AF316145_1 (31.94% identity, 47.57% similarity) AAA22332.1 (29.07%identity, 47.28% similarity) ANN35812.1 (28.16% identity, 40.53%similarity) APG03715 45, 50, 55, 60, 65, APG00696 70, 75, 80, 85, 90,(92.19% identity, 95, 96, 97, 98, 99 93.13% similarity) APG01882 (92.19%identity, 93.13% similarity) APG06465 (91.88% identity, 93.75%similarity) APG04483 (91.88% identity, 93.44% similarity) APG08138(90.63% identity, 92.50% similarity) AGP18056.1 (24.77% identity, 39.88%similarity) US20120278954A1_26 (24.35% identity, 41.16% similarity)Cry33Aa1 (24.11% identity, 40.18% similarity) APG03747 50, 55, 60, 65,70, APG04793 75, 80, 85, 90, 95, (88.39% identity, 96, 97, 98, 99 93.63%similarity) APG03686 (87.64% identity, 94.01% similarity) APG07676(86.14% identity, 93.63% similarity) WP_033694890.1 (33.43% identity,46.24% similarity) AF316145_1 (30.21% identity, 47.22% similarity)AAA22332.1 (28.53% identity, 44.51% similarity) ANN35812.1 (27.36%identity, 40.92% similarity) APG03760 80, 85, 90, 95, 96, APG00224 97,98, 99 (78.21% identity, 82.96% similarity) AGA40030.1 (66.39% identity,78.71% similarity) APG05634 (65.24% identity, 75.78% similarity)APG00609 (64.96% identity, 75.50% similarity) APG00846 (54.8% identity,70.90% similarity) APG00513 (54.24% identity, 71.19% similarity)APG02280 (53.30% identity, 71.06% similarity) US_8829279_B2-27 (50.43%identity, 66.67% similarity) CAA67205.1 (44.48% identity, 58.92%similarity) AGA40032.1 (40.22% identity, 55.10% similarity) APG03831 96,97, 98, 99 APG03238 (97.00% identity, 98.29% similarity) APG04686(96.79% identity, 98.29% similarity) APG08151 (96.36% identity, 98.29%similarity) APG06338 (95.93% identity, 97.64% similarity) KIF65774.1(95.50% identity, 95.72% similarity) KHA73845.1 (70.45% identity, 80.09%similarity) WP_042557199.1 (59.70% identity, 70.36% similarity)WP_064117037.1 (57.66% identity, 68.94% similarity) APG03867 55, 60, 65,70, 75, APG00143 80, 85, 90, 95, 96, (68.63% identity, 97, 98, 99 81.99%similarity) APG00441 (67.29% identity, 80.69% similarity) APG00444(66.67% identity, 80.06% similarity) APG00833 (65.95% identity, 80.98%similarity) APG07682 (63.86% identity, 75.30% similarity) WP_006918908.1(31.8% identity, 50.46% similarity) APG04067 50, 55, 60, 65, 70,APG07114 75, 80, 85, 90, 95, (69.07% identity, 96, 97, 98, 99 83.18%similarity) APG02531 (67.84% identity, 81.29% similarity) CA_2844913-114(30.71% identity, 47.01% similarity) CA_2844913-111_1 (29.18% identity,46.15% similarity) APG04099 65, 70, 75, 80, 85, E6I1C3_ENTFL 90, 95, 96,97, 98, (42.04% identity, 99 58.86% similarity) WP_025188414.1 (41.16%identity, 57.68% similarity) R2T0I9_9ENTE (37.69% identity, 60.24%similarity) R2N0B3_ENTMU (31.56% identity, 54.75% similarity) APG0415245, 50, 55, 60, 65, WP_044307385.1 70, 75, 80, 85, 90, (28.90% identity,95, 96, 97, 98, 99 42.81% similarity) WP_048536363.1 (26.37% identity,42.70% similarity) WP_048536324.1 (25.85% identity, 39.82% similarity)AGA40057.1 (25.70% identity, 37.29% similarity) APG04450 95, 96, 97, 98,99 APG01507 (85.58% identity, 92.63% similarity) WP_016132978.1 (84.94%identity, 93.27% similarity) WP_016113347.1 (84.62% identity, 92.95%similarity) WP_016131662.1 (84.29% identity, 92.95% similarity) APG06589(80.29% identity, 85.51% similarity) APG02921 (78.27% identity, 87.86%similarity) APG06989 (77.32% identity, 88.18% similarity) APG04483 45,50, 55, 60, 65, APG06465 70, 75, 80, 85, 90, (98.69% identity, 95, 96,97, 98, 99 99.02% similarity) APG00696 (96.39% identity, 97.38%similarity) APG01882 (95.75% identity, 97.06% similarity) APG08138(95.08% identity, 97.38% similarity) APG03715 (91.88% identity, 93.44%similarity) Cry33Aa1 (25.93% identity, 42.59% similarity) APG04686 95,96, 97, 98, 99 APG03238 (98.50% identity, 99.14% similarity) APG08151(98.50% identity, 99.14% similarity) APG06338 (97.43% identity, 98.50%similarity) APG03831 (96.79% identity, 98.29% similarity) KIF65774.1(93.15% identity, 94.65% similarity) KHA73845.1 (70.02% identity, 80.09%similarity) WP_042557199.1 (58.85% identity, 69.51% similarity)WP_064117037.1 (56.78% identity, 67.58% similarity) APG04721 40, 45, 50,55, 60, WP_036155538.1 65, 70, 75, 80, 85, (24.16% identity, 90, 95, 96,97, 98, 38.10% similarity) 99 WP_049738340.1 (21.57% identity, 36.93%similarity) WP_025143599.1 (20.93% identity, 36.07% similarity)EXX63903.1 (18.23% identity, 35.20% similarity) APG04778 35, 40, 45, 50,55, AGA40063.1 60, 65, 70, 75, 80, (20.94% identity, 85, 90, 95, 96, 97,30.00% similarity) 98, 99 ADE27985.1 (19.94% identity, 26.19%similarity) WP_044797748.1 (18.81% identity, 27.59% similarity) Cry60Aa2(16.36% identity, 22.22% similarity) APG04793 50, 55, 60, 65, 70,APG03686 75, 80, 85, 90, 95, (96.25% identity, 96, 97, 98, 99 98.13%similarity) APG07676 (90.64% identity, 94.38% similarity) APG03747(88.39% identity, 93.63% similarity) APG00014 (51.11% identity, 66.67%similarity) WP_033694890.1 (34.35% identity, 46.26% similarity)AF316145_1 (31.25% identity, 47.22% similarity) AAA22332.1 (28.43%identity, 46.96% similarity) ANN35812.1 (27.67% identity, 40.29%similarity) APG04925 45, 50, 55, 60, 65, V9Z0X8_BURPE 70, 75, 80, 85,90, (26.37% identity, 95, 96, 97, 98, 99 42.08% similarity)WP_009971549.1 (26.30% identity, 41.78% similarity) WP_057050573.1(26.28% identity, 42.46% similarity) WP_038779659.1 (26.24% identity,42.03% similarity) APG05213 99 WP_016986342.1 (97.59% identity, 98.91%similarity) WP_064117037.1 (67.40% identity, 78.34% similarity)KIP93183.1 (66.30% identity, 77.24% similarity) KPG83347.1 (66.30%identity, 77.24% similarity) APG03831 (53.49% identity, 68.08%similarity) APG04686 (52.64% identity, 67.23% similarity) APG08151(52.64% identity, 67.02% similarity) APG06338 (52.13% identity, 67.23%similarity) APG05372 55, 60, 65, 70, 75, APG00293 80, 85, 90, 95, 96,(61.42% identity, 97, 98, 99 68.77% similarity) APG00170 (55.53%identity, 62.56% similarity) EJQ15321.1 (35.80% identity, 50.31%similarity) C3ICE4_BACTU (33.72% identity, 47.26% similarity) SCA97082.1(33.72% identity, 47.26% similarity) J8Y0J8_BACCE (33.14% identity,46.97% similarity) APG05500 60, 65, 70, 75, 80, CA_2868815-146 85, 90,95, 96, 97, (42.19% identity, 98, 99 59.38% similarity) US_9322033_B2-51(41.67% identity, 58.64% similarity) US_9322033_B2-55 (41.67% identity,58.64% similarity) Cry51Aa1 (41.28% identity, 57.80% similarity)APG05553 80, 85, 90, 95, 96, WP_035054261.1 97, 98, 99 (69.23% identity,77.95% similarity) WP_035054034.1 (33.89% identity, 52.18% similarity)WP_025141994.1 (26.66% identity, 43.30% similarity) XP_005716346.1(22.49% identity, 38.55% similarity) APG05634 75, 80, 85, 90, 95,APG00609 96, 97, 98, 99 (79.11% identity, 82.73% similarity) APG00224(65.72% identity, 76.77% similarity) APG03760 (65.24% identity, 75.78%similarity) AGA40030.1 (63.03% identity, 74.23% similarity) APG02280(54.83% identity, 70.17% similarity) APG00846 (54.21% identity, 69.38%similarity) APG00513 (54.19% identity, 70.39% similarity)US_8829279_B2-27 (48.17% identity, 62.82% similarity) APG05706 96, 97,98, 99 BAI44028.1 (91.54% identity, 95.05% similarity) BAI44022.1(23.96% identity, 38.02% similarity) AGO57767.1 (23.63% identity, 37.58%similarity) Cry2Ah2 (18.97% identity, 35.59% similarity) APG06001 40,45, 50, 55, 60, WP_006918908.1 65, 70, 75, 80, 85, (22.78% identity, 90,95, 96, 97, 98, 39.44% similarity) 99 APG06281 25, 30, 35, 40, 45,ACU24782.1 50, 55, 60, 65, 70, (14.14% identity, 75, 80, 85, 90, 95,19.41% similarity) 96, 97, 98, 99 CA_2844913-86 (14.14% identity, 19.41%similarity) Cry60Ba1 (13.89% identity, 20.68% similarity) APG06324 95,96, 97, 98, 99 WP_003304158.1 (85.84% identity, 93.48% similarity)WP_003305942.1 (83.00% identity, 91.78% similarity) AGP18070.1 (76.10%identity, 84.16% similarity) US_8829279_B2-6 (59.33% identity, 72.14%similarity) APG06338 95, 96, 97, 98, 99 APG08151 (97.64% identity,98.50% similarity) APG04686 (97.43% identity, 98.50% similarity)APG03238 (96.79% identity, 98.07% similarity) APG03831 (95.93% identity,97.64% similarity) KIF65774.1 (92.29% identity, 94.00% similarity)KHA73845.1 (70.66% identity, 79.87% similarity) WP_042557199.1 (59.28%identity, 69.72% similarity) WP_064117037.1 (56.81% identity, 68.30%similarity) APG06381 45, 50, 55, 60, 65, APG02531 70, 75, 80, 85, 90,(23.90% identity, 95, 96, 97, 98, 99 42.31% similarity)US_2013_0227743_A1-102 (23.68% identity, 34.76% similarity) APG07114(23.35% identity, 41.48% similarity) US_2013_0227743_A1-114 (22.67%identity, 38.67% similarity) APG06465 45, 50, 55, 60, 65, APG04483 70,75, 80, 85, 90, (98.69% identity, 95, 96, 97, 98, 99 99.02% similarity)APG00696 (96.72% identity, 97.70% similarity) APG01882 (95.75% identity,97.39% similarity) APG08138 (95.74% identity, 97.70% similarity)APG03715 (91.88% identity, 93.75% similarity) Cry33Aa1 (25.93% identity,41.98% similarity) APG06501 45, 50, 55, 60, 65, WP_003209347.1 70, 75,80, 85, 90, (26.65% identity, 95, 96, 97, 98, 99 44.99% similarity)WP_018767932.1 (25.79% identity, 44.41% similarity) WP_035437465.1(24.57% identity, 43.14% similarity) Cry55Aa1 (22.16% identity, 35.31%similarity) APG06589 85, 90, 95, 96, 97, APG01507 98, 99 (84.84%identity, 87.17% similarity) APG04450 (80.29% identity, 85.51%similarity) WP_016132978.1 (76.68% identity, 83.97% similarity)WP_016113347.1 (76.38% identity, 83.67% similarity) WP_016131662.1(75.80% identity, 83.67% similarity) APG02921 (73.91% identity, 80.87%similarity) APG06989 (73.04% identity, 80.87% similarity) Cry55Aa1(19.81% identity, 35.14% similarity) APG06676 85, 90, 95, 96, 97,WP_025688444.1 98, 99 (70.95% identity, 81.42% similarity) APG06894 95,96, 97, 98, 99 WP_062759953.1 (87.50% identity, 93.23% similarity)WP_055639819.1 (87.50% identity, 92.71% similarity) WP_056557960.1(86.98% identity, 93.23% similarity) APG00134 (86.98% identity, 92.19%similarity) APG00198 (86.98% identity, 92.19% similarity) APG08780(72.16% identity, 84.54% similarity) APG00945 (70.41% identity, 84.69%similarity) APG00108 (62.41% identity, 66.54% similarity) APG00138(51.38% identity, 54.46% similarity) APG06989 95, 96, 97, 98, 99APG02921 (97.76% identity, 98.4% similarity) APG01536 (88.46% identity,93.59% similarity) WP_025150761.1 (88.14% identity, 93.27% similarity)WP_016131662.1 (81.73% identity, 89.42% similarity) WP_016132978.1(81.41% identity, 89.74% similarity) APG01507 (80.45% identity, 89.74%similarity) APG06589 (73.04% identity, 80.87% similarity) APG06997 60,65, 70, 75, 80, AGA40057.1 85, 90, 95, 96, 97, (38.79% identity, 98, 9957.45% similarity) WP_048536324.1 (34.83% identity, 48.43% similarity)WP_017762619.1 (33.80% identity, 50.93% similarity) WP_048536362.1(33.70% identity, 51.57% similarity) APG07002 45, 50, 55, 60, 65,WP_016113505.1 70, 75, 80, 85, 90, (27.11% identity, 95, 96, 97, 98, 9942.27% similarity) WP_002193657.1 (26.59% identity, 42.49% similarity)WP_006097189.1 (26.39% identity, 43.70% similarity) WP_016113347.1(26.29% identity, 42.00% similarity) APG07020 70, 75, 80, 85, 90,APG00330 95, 96, 97, 98, 99 (59.07% identity, 70.46% similarity)WP_059525806.1 (54.38% identity, 65.55% similarity) WP_059692427.1(54.38% identity, 65.36% similarity) WP_059741244.1 (54.09% identity,67.66% similarity) APG07114 50, 55, 60, 65, 70, APG02531 75, 80, 85, 90,95, (94.10% identity, 96, 97, 98, 99 95.28% similarity) APG04067 (69.07%identity, 83.18% similarity) CA_2844913-114 (29.19% identity, 46.76%similarity) US20130227743A1_200 (16.92% identity, 23.93% similarity)AGA40058.1 (13.6% identity, 20.73% similarity) CA_2844913-111_1 (29.49%identity, 43.85% similarity) APG07220 45, 50, 55, 60, 65, WP_016873057.170, 75, 80, 85, 90, (23.76% identity, 95, 96, 97, 98, 99 44.36%similarity) G8LV29_CLOCD (20.59% identity, 33.82% similarity)WP_016873055.1 (16.47% identity, 29.52% similarity) APG07224 60, 65, 70,75, 80, WP_044307385.1 85, 90, 95, 96, 97, (40.78% identity, 98, 9955.5% similarity) AGA40057.1 (36.88% identity, 51.37% similarity)WP_048536362.1 (36.11% identity, 49.14% similarity) WP_017762616.1(29.27% identity, 43.35% similarity) APG07444 50, 55, 60, 65, 70,WP_065397681.1 75, 80, 85, 90, 95, (29.81% identity, 96, 97, 98, 9945.96% similarity) WP_053730552.1 (28.93% identity, 41.57% similarity)CTX_PSEAI (27.69% identity, 43.08% similarity) WP_037683010.1 (27.63%identity, 41.74% similarity) APG07445 35, 40, 45, 50, 55,US20130227743A1_200 60, 65, 70, 75, 80, (18.62% identity, 85, 90, 95,96, 97, 30.2% similarity) 98, 99 AGA40057.1 (17.69% identity, 30.91%similarity) AGA40058.1 (17.50% identity, 30.18% similarity)US20130227743A1_152 (15.31% identity, 27.12% similarity) APG07470 20,25, 30, 35, 40, WP_044306828.1 45, 50, 55, 60, 65, (12.09% identity, 70,75, 80, 85, 90, 19.88% similarity) 95, 96, 97, 98, 99 AGA40058.1 (11.54%identity, 17.50% similarity) WP_044306756.1 (10.76% identity, 18.57%similarity) WP_048536362.1 (10.63% identity, 16.47% similarity) APG0763997, 98, 99 J8YPM2_BACCE (94.88% identity, 96.93% similarity)CA_2844913-100 (94.54% identity, 96.93% similarity) APG00155 (78.79%identity, 85.19% similarity) APG00006 (78.69% identity, 84.59%similarity) APG00107 (77.36% identity, 87.84% similarity) APG00749(77.00% identity, 84.33% similarity) APG00201 (75.08% identity, 82.85%similarity) APG00847 (74.43% identity, 83.17% similarity) APG00589(71.48% identity, 81.88% similarity) APG01451 (67.00% identity, 72.39%similarity) APG00955 (65.48% identity, 74.52% similarity) CA_2844913-60(41.45% identity, 46.38% similarity) WP_051437016.1 (24.23% identity,36.50% similarity) APG07676 50, 55, 60, 65, 70, APG04793 75, 80, 85, 90,95, (90.64% identity, 96, 97, 98, 99 94.38% similarity) APG03686 (89.14%identity, 94.38% similarity) APG03747 (86.14% identity, 93.63%similarity) APG00014 (50.74% identity, 67.78% similarity) WP_033694890.1(34.07% identity, 45.98% similarity) US_2012_0278954_A1-22 (32.63%identity, 49.47% similarity) AF316145_1 (30.90% identity, 47.92%similarity) AAA22332.1 (30.15% identity, 46.77% similarity) APG07682 55,60, 65, 70, 75, APG00441 80, 85, 90, 95, 96, (67.27% identity, 97, 98,99 80.30% similarity) APG00143 (66.87% identity, 79.70% similarity)APG00444 (65.77% identity, 78.57% similarity) APG03867 (63.86% identity,75.30% similarity) APG00833 (63.66% identity, 77.18% similarity)WP_006918908.1 (32.33% identity, 50.15% similarity) APG07738 40, 45, 50,55, 60, APG02633 65, 70, 75, 80, 85, (26.92% identity, 90, 95, 96, 97,98, 41.99% similarity) 99 US_2015_0047076_A1-12 (24.09% identity, 37.20%similarity) US_8461421_B2-102 (23.84% identity, 38.08% similarity)Cry45Aa1 (23.62% identity, 36.81% similarity) Cry23Aa1 (23.25% identity,37.26% similarity) APG09455 (23.10% identity, 40.64% similarity)APG07780 35, 40, 45, 50, 55, APG01273 60, 65, 70, 75, 80, (70.07%identity, 85, 90, 95, 96, 97, 82.04% similarity) 98, 99 WP_033694890.1(21.04% identity, 31.95% similarity) ANN35812.1 (15.78% identity, 25.52%similarity) APG07954 90, 95, 96, 97, 98, 99 APG00965 (86.42% identity,90.22% similarity) WP_001087511.1 (79.74% identity, 87.17% similarity)CA_2844913-12 (79.59% identity, 86.73% similarity) APG00152 (76.97%identity, 86.15% similarity) EEM92570.1 (72.74% identity, 80.32%similarity) APG00084 (60.9% identity, 71.45% similarity) APG08029 50,55, 60, 65, 70, WP_033694890.1 75, 80, 85, 90, 95, (29.38% identity, 96,97, 98, 99 46.17% similarity) ANN35812.1 (26.89% identity, 40.67%similarity) AAA22332.1 (23.23% identity, 35.99% similarity) APG08138 45,50, 55, 60, 65, APG01882 70, 75, 80, 85, 90, (96.73% identity, 95, 96,97, 98, 99 97.71% similarity) APG06465 (95.74% identity, 97.70%similarity) APG00696 (95.74% identity, 96.72% similarity) APG04483(95.08% identity, 97.38% similarity) APG03715 (90.63% identity, 92.50%similarity) AGP18056.1 (25.23% identity, 40.31% similarity) US +2012_0278954_A1-26 (25.15% identity, 43.33% similarity)US_2015_0047076_A1-6 (25.00% identity, 44.62% similarity) Cry45Aa(24.38% identity, 41.36% similarity) APG08151 95, 96, 97, 98, 99APG04686 (98.50% identity, 99.14% similarity) APG03238 (97.86% identity,98.72% similarity) APG06338 (97.64% identity, 98.50% similarity)APG03831 (96.36% identity, 98.29% similarity) KIF65774.1 (92.72%identity, 94.65% similarity) KHA73845.1 (69.81% identity, 79.44%similarity) WP_042557199.1 (58.64% identity, 69.51% similarity)WP_064117037.1 (56.57% identity, 67.58% similarity) APG08509 85, 90, 95,96, 97, A0A030UY73_PSEAI 98, 99 (74.05% identity, 82.59% similarity)WP_039843228.1 (70.89% identity, 80.70% similarity) WP_058177796.1(70.89% identity, 80.38% similarity) CTX_PSEAI (70.57% identity, 80.7%similarity) APG08607 45, 50, 55, 60, 65, WP_048536362.1 70, 75, 80, 85,90, (30.78% identity, 95, 96, 97, 98, 99 43.82% similarity)WP_017762581.1 (29.11% identity, 40.03% similarity) AGP17992.1 (29.10%identity, 39.89% similarity) WP_044306759.1 (29.10% identity, 39.89%similarity) APG08628 25, 30, 35, 40, 45, WP_044306756.1 50, 55, 60, 65,70, (20.55% identity, 75, 80, 85, 90, 95, 23.09% similarity) 96, 97, 98,99 APG08780 95, 96, 97, 98, 99 WP_046912431.1 (91.75% identity, 94.33%similarity) APG00134 (76.80% identity, 87.63% similarity) KUM91086.1(75.77% identity, 83.51% similarity) WP_055599179.1 (75.26% identity,87.11% similarity) APG00945 (74.87% identity, 81.91% similarity)APG06894 (72.16% identity, 84.54% similarity) APG00327 (68.22% identity,70.93% similarity) APG00108 (54.48% identity, 62.69% similarity)APG08794 50, 55, 60, 65, 70, WP_065486138.1 75, 80, 85, 90, 95, (29.26%identity, 96, 97, 98, 99 47.16% similarity) WP_065487080.1 (28.98%identity, 46.88% similarity) C3ICE4_BACTU (28.94% identity, 46.42%similarity) J8Y0J8_BACCE (28.94% identity, 46.42% similarity) APG0905575, 80, 85, 90, 95, APG00569 96, 97, 98, 99 (82.74% identity, 89.29%similarity) APG00938 (82.14% identity, 89.29% similarity) APG00563(76.49% identity, 84.82% similarity) APG00794 (72.70% identity, 83.68%similarity) APG00387 (63.45% identity, 76.32% similarity) C3GC23_BACTU(60.36% identity, 73.08% similarity) AGA40045.1 (59.35% identity, 73.29%similarity) ANN35739.1 (58.58% identity, 73.96% similarity)US_2013_0227743_A1-102 (57.99% identity, 73.67% similarity) APG09096 65,70, 75, 80, 85, APG00490 - 90, 95, 96, 97, 98, US_2016_0177333_A1-143 99(44.69% identity, 61.61% similarity) APG00490.1 - US_2016_0177333_A1-144(44.69% identity, 61.61% similarity) WP_017154368.1 (44.60% identity,61.69% similarity) Cry22Bb1 (34.74% identity, 47.52% similarity)APG09376 95, 96, 97, 98, 99 APG00116 (91.19% identity, 95.22%similarity) AFM37573.1 (90.58% identity, 94.77% similarity) AGP18043.1(90.18% identity, 94.35% similarity) AGV55020.1 (83.56% identity, 91.18%similarity) APG00153 (53.95% identity, 64.42% similarity) Cry53Aa1(37.92% identity, 53.23% similarity) APG09455 40, 45, 50, 55, 60,APG00598 65, 70, 75, 80, 85, (36.07% identity, 90, 95, 96, 97, 98,52.20% similarity) 99 APG02633 (29.04% identity, 42.81% similarity)APG00236 (28.04% identity, 47.35% similarity) Cry23Aa1 (23.90% identity,39.94% similarity) APG09642 50, 55, 60, 65, 70, R2N0B3_ENTMU 75, 80, 85,90, 95, (31.09% identity, 96, 97, 98, 99 48.18% similarity) E6I1C3_ENTFL(29.50% identity, 48.67% similarity) WP_025188414.1 (28.41% identity,46.88% similarity) WP_062805258.1 (27.76% identity, 44.18% similarity)APG09659 25, 30, 35, 40, 45, WP_048536363.1 50, 55, 60, 65, 70, (12.68%identity, 75, 80, 85, 90, 95, 18.21% similarity) 96, 97, 98, 99AGA40058.1 (12.12% identity, 20.00% similarity) US_2013_0227743_A1-152(11.88% identity, 17.55% similarity) WP_048536362.1 (11.07% identity,16.52% similarity) APG09717 95, 96, 97, 98, 99 APG00456 (95.10%identity, 98.04% similarity) WP_003203846.1 (92.65% identity, 94.61%similarity) WP_033798332.1 (92.65% identity, 94.61% similarity)WP_040119538.1 (92.65% identity, 94.61% similarity) APG09726 55, 60, 65,70, 75, ANS51604.1 80, 85, 90, 95, 96, (45.60% identity, 97, 98, 9954.67% similarity) US_2016_0017363_A1-60 (44.53% identity, 53.60%similarity) US_2016_0017363_A1-61 (44.27% identity, 53.33% similarity)APG09735 50, 55, 60, 65, 70, WP_025188414.1 75, 80, 85, 90, 95, (32.05%identity, 96, 97, 98, 99 48.97% similarity) E6I1C3_ENTFL (31.28%identity, 47.18% similarity) R2N0B3_ENTMU (29.50% identity, 48.56%similarity) US_8461415_B2-50 (28.39% identity, 44.27% similarity)APG00743 99 WP_044444098.1 (96.04% identity, 98.35% similarity) APG02555(92.74% identity, 97.03% similarity) APG08085 (90.76% identity, 96.37%similarity) APG04643 (90.12% identity, 91.98% similarity) WP_060749709.1(87.83% identity, 93.09% similarity) SFC26517.1 (80.52% identity, 89.94%similarity) APG04226 (53.65% identity, 70.48% similarity) Cry55Aa1(20.63% identity, 36.81% similarity) APG01705 95, 96, 97, 98, 99APG03368 (99.35% identity, 100.0% similarity) APG01507 (95.81% identity,98.06% similarity) APG04224 (89.8% identity, 90.09% similarity) APG06589(87.76% identity, 88.92% similarity) WP_016132978.1 (84.52% identity,92.9% similarity) WP_016113347.1 (84.19% identity, 92.58% similarity)WP_016131662.1 (83.23% identity, 92.9% similarity) Cry55Aa1 (19.46%identity, 33.99% similarity) APG01989 100 WP_026594351.1 (100.0%identity, 100.0% similarity) SDY81229.1 (99.51% identity, 99.51%similarity) WP_003203846.1 (94.61% identity, 96.57% similarity) APG00456(92.65% identity, 95.1% similarity) APG09717 (92.16% identity, 94.12%similarity) Cyt2Ca1 (24.29% identity, 41.7% similarity) APG02245 99WP_000586614.1 (96.44% identity, 98.02% similarity) WP_065230018.1(95.26% identity, 97.23% similarity) WP_016082893.1 (94.47% identity,97.63% similarity) APG09256 (93.7% identity, 96.06% similarity) APG01269(92.91% identity, 95.28% similarity) APG01103 (91.7% identity, 97.23%similarity) APG02768 (91.7% identity, 96.84% similarity) Cry46Ab (28.01%identity, 42.35% similarity) APG02279 95, 96, 97, 98, 99 APG08241(91.39% identity, 92.72% similarity) APG08718 (91.06% identity, 93.38%similarity) J8YPM2_BACCE (91.06% identity, 93.38% similarity) APG07639(88.08% identity, 91.06% similarity) APG00006 (87.21% identity, 92.13%similarity) US_2016_0304898_A1-9 (87.21% identity, 92.13% similarity)US20130227743A1_100 (87.09% identity, 91.39% similarity) APG00201(79.02% identity, 87.21% similarity) APG00566 (78.76% identity, 88.24%similarity) APG00847 (78.36% identity, 87.21% similarity) APG00749(77.7% identity, 86.56% similarity) APG00107 (75.74% identity, 85.25%similarity) US20130227743A1_60 (44.7% identity, 49.67% similarity)APG02429 100 WP_044585299.1 (96.84% identity, 99.21% similarity)WP_000586617.1 (96.84% identity, 98.81% similarity) APG01103 (95.65%identity, 98.81% similarity) WP_000586615.1 (95.65% identity, 98.42%similarity) APG09842 (95.65% identity, 98.02% similarity) APG02768(93.68% identity, 98.42% similarity) APG00737 (93.68% identity, 96.44%similarity) Cry46Ab (30.16% identity, 43.93% similarity) APG02674 98, 99WP_061688274.1 (96.26% identity, 97.13% similarity) APG01420 (95.4%identity, 97.13% similarity) WP_050595402.1 (92.53% identity, 93.1%similarity) WP_023524027.1 (50.42% identity, 64.82% similarity) Cry55Aa1(20.95% identity, 34.16% similarity) APG02768 100 WP_061667036.1 (96.44%identity, 98.42% similarity) ADQ73630.1 (95.65% identity, 98.02%similarity) WP_044585299.1 (95.26% identity, 99.21% similarity) APG02429(93.68% identity, 98.42% similarity) APG09842 (92.49% identity, 98.02%similarity) APG00737 (92.49% identity, 96.05% similarity) APG01103(92.09% identity, 98.02% similarity) Cry46Ab (28.85% identity, 43.28%similarity) APG03040 100 WP_064474064.1 (98.81% identity, 99.21%similarity) APG01150 (98.02% identity, 98.42% similarity) WP_061530406.1(98.02% identity, 98.42% similarity) APG00788 (97.23% identity, 98.02%similarity) SCA96918.1 (96.84% identity, 97.63% similarity) APG00737(83.4% identity, 90.12% similarity) APG02429 (83.0% identity, 89.72%similarity) Cry46Ab (26.28% identity, 40.71% similarity) APG03185 95,96, 97, 98, 99 APG07574 (94.22% identity, 96.66% similarity) APG02518(92.71% identity, 95.74% similarity) WP_047426867.1 (87.58% identity,93.03% similarity) WP_062673074.1 (87.27% identity, 93.94% similarity)WP_034734550.1 (77.64% identity, 89.73% similarity) WP_027372020.1(76.74% identity, 89.12% similarity) APG03114 (76.67% identity, 88.79%similarity) APG03217 45, 50, 55, 60, 65, APG08029 70, 75, 80, 85, 90,(97.42% identity, 95, 96, 97, 98, 99 98.45% similarity) ANN35812.1(25.16% identity, 38.73% similarity) WP_033694890.1 (24.71% identity,40.5% similarity) US_2016_0304898_A1-24 (23.74% identity, 36.11%similarity) AAA22332.1 (23.18% identity, 36.36% similarity) APG03368 95,96, 97, 98, 99 APG01705 (99.35% identity, 100.0% similarity) APG01507(95.81% identity, 98.06% similarity) APG04224 (89.8% identity, 90.09%similarity) APG06589 (87.76% identity, 88.92% similarity) WP_016132978.1(85.16% identity, 92.9% similarity) WP_016113347.1 (84.84% identity,92.58% similarity) WP_016131662.1 (83.87% identity, 92.9% similarity)Cry55Aa1 (19.21% identity, 33.99% similarity) APG03662 75, 80, 85, 90,95, APG02038 96, 97, 98, 99 (91.03% identity, 93.59% similarity)US_8829279_B2-25 (63.28% identity, 74.03% similarity) APG00661 (63.1%identity, 73.81% similarity) AGA40029.1 (61.7% identity, 73.86%similarity) US_8829279_B2-35 (59.88% identity, 72.95% similarity)APG00528 (55.76% identity, 63.53% similarity) AGA40031.1 (51.25%identity, 68.44% similarity) APG04224 85, 90, 95, 96, 97, APG06589 98,99 (97.38% identity, 98.25% similarity) APG01705 (89.8% identity, 90.09%similarity) APG03368 (89.8% identity, 90.09% similarity) APG01507(86.59% identity, 88.34% similarity) WP_016113347.1 (76.68% identity,83.97% similarity) WP_016132978.1 (76.38% identity, 83.67% similarity)WP_016131662.1 (75.22% identity, 83.67% similarity) Cry55Aa1 (19.81%identity, 34.91% similarity) APG04226 95, 96, 97, 98, 99 APG05969(96.47% identity, 99.04% similarity) APG04485 (93.91% identity, 97.76%similarity) APG02921 (89.42% identity, 94.23% similarity) APG08990(89.1% identity, 94.55% similarity) WP_025150761.1 (86.22% identity,92.31% similarity) SFS68380.1 (84.39% identity, 92.04% similarity)WP_016131662.1 (82.37% identity, 90.71% similarity) Cry55Aa1 (17.6%identity, 34.47% similarity) APG04485 95, 96, 97, 98, 99 APG04226(93.91% identity, 97.76% similarity) APG05969 (93.59% identity, 98.08%similarity) APG02921 (88.78% identity, 94.23% similarity) APG08990(88.46% identity, 94.87% similarity) SFS68380.1 (85.99% identity, 92.99%similarity) WP_025150761.1 (85.9% identity, 91.67% similarity)WP_016131662.1 (83.01% identity, 90.71% similarity) Cry55Aa1 (17.87%identity, 33.09% similarity) APG04643 95, 96, 97, 98, 99 WP_044444098.1(92.59% identity, 93.52% similarity) APG00743 (90.12% identity, 91.98%similarity) APG02555 (87.65% identity, 91.67% similarity) APG08085(84.26% identity, 90.12% similarity) WP_060749709.1 (82.15% identity,87.69% similarity) SFC26517.1 (75.38% identity, 84.5% similarity)APG04226 (50.3% identity, 66.67% similarity) Cry55Aa1 (21.52% identity,37.22% similarity) APG05660 98, 99 US_2016_0311864_A1-233 (96.1%identity, 97.9% similarity) APG00116 (95.67% identity, 97.46%similarity) US_2016_0311864_A1-25 (95.67% identity, 97.46% similarity)APG09376 (95.05% identity, 97.45% similarity) US_2016_0311864_A1-234(92.35% identity, 94.0% similarity) APG06528 (90.9% identity, 94.78%similarity) APG00153 (53.56% identity, 64.17% similarity) Cry53Aa1(38.23% identity, 54.07% similarity) APG05969 95, 96, 97, 98, 99APG04226 (96.47% identity, 99.04% similarity) APG04485 (93.59% identity,98.08% similarity) APG08990 (90.06% identity, 94.87% similarity)APG02921 (89.1% identity, 94.23% similarity) WP_025150761.1 (86.22%identity, 91.99% similarity) SFS68380.1 (85.67% identity, 92.36%similarity) WP_016132978.1 (83.33% identity, 91.35% similarity) Cry55Aa1(18.38% identity, 33.33% similarity) APG06528 98, 99 APG00116 (94.33%identity, 97.31% similarity) US_2016_0311864_A1-25 (94.33% identity,97.31% similarity) CA_2729294-29_1 (94.05% identity, 97.17% similarity)AGP18043.1 (93.9% identity, 97.17% similarity) APG09376 (91.04%identity, 95.22% similarity) APG05660 (90.9% identity, 94.78%similarity) APG00153 (53.93% identity, 64.48% similarity) Cry53Aa1(39.94% identity, 54.29% similarity) APG07049 95, 96, 97, 98, 99APG02921 (93.59% identity, 96.47% similarity) APG06989 (92.63% identity,96.79% similarity) APG08990 (91.67% identity, 95.83% similarity)APG04226 (89.42% identity, 93.27% similarity) WP_025150761.1 (88.46%identity, 93.27% similarity) SFS68380.1 (85.35% identity, 91.4%similarity) WP_016131662.1 (82.05% identity, 88.46% similarity) Cry55Aa1(17.24% identity, 34.24% similarity) APG07574 95, 96, 97, 98, 99APG03185 (94.22% identity, 96.66% similarity) APG02518 (93.62% identity,96.05% similarity) WP_047426867.1 (88.18% identity, 93.64% similarity)WP_062673074.1 (87.88% identity, 93.03% similarity) WP_034734550.1(77.58% identity, 89.7% similarity) APG03114 (77.27% identity, 89.09%similarity) WP_027372020.1 (76.67% identity, 89.09% similarity) APG0804340, 45, 50, 55, 60, APG07738 65, 70, 75, 80, 85, (96.95% identity, 90,95, 96, 97, 98, 99 97.97% similarity) APG08085 97, 98, 99 APG02555(95.05% identity, 97.69% similarity) WP_044444098.1 (91.09% identity,96.37% similarity) APG00743 (90.76% identity, 96.37% similarity)WP_060749709.1 (86.51% identity, 94.08% similarity) APG04643 (84.26%identity, 90.12% similarity) SFC26517.1 (80.84% identity, 89.94%similarity) APG04226 (54.92% identity, 70.16% similarity) Cry55Aa1(19.31% identity, 35.98% similarity) APG08225 98, 99 APG01022 (99.46%identity, 99.46% similarity) APG02400 (99.19% identity, 99.46%similarity) US_2016_0311864_A1-267 (97.58% identity, 97.85% similarity)APG00426 (97.32% identity, 98.12% similarity) APG00891 (92.56% identity,93.33% similarity) APG00227 (90.75% identity, 91.0% similarity)US_2016_0311864_A1-83 (90.75% identity, 91.0% similarity)US_2016_0311864_A1-268 (89.78% identity, 89.78% similarity) APG00517(80.97% identity, 82.57% similarity) APG00158 (64.46% identity, 72.68%similarity) WP_071770709.1 (58.13% identity, 69.33% similarity) APG01508(56.91% identity, 69.41% similarity) APG08241 97, 98, 99 APG08718(95.22% identity, 96.93% similarity) J8YPM2_BACCE (95.22% identity,96.93% similarity) APG07639 (91.81% identity, 94.54% similarity)APG02279 (91.39% identity, 92.72% similarity) US20130227743A1_100(90.78% identity, 94.54% similarity) APG00107 (79.39% identity, 89.86%similarity) US_2016_0311864_A1-21 (79.39% identity, 89.86% similarity)APG00006 (79.34% identity, 85.57% similarity) APG00930 (78.72% identity,88.85% similarity) APG00749 (77.67% identity, 85.67% similarity)APG00847 (76.97% identity, 85.53% similarity) APG00201 (76.97% identity,85.2% similarity) US20130227743A1_60 (43.05% identity, 48.34%similarity) APG08411 95, 96, 97, 98, 99 APG00989 (92.1% identity, 95.74%similarity) AOB42285.1 (86.02% identity, 92.71% similarity) APG00309(79.76% identity, 87.01% similarity) C3HSG6_BACTU (66.77% identity,80.36% similarity) WP_018673409.1 (62.28% identity, 75.75% similarity)APG00585 (61.38% identity, 74.85% similarity) APG00427 (60.98% identity,75.3% similarity) US_2016_0311864_A1-178 (60.98% identity, 75.3%similarity) APG00851 (60.37% identity, 75.0% similarity) APG08718 100WP_000963933.1 (100.0% identity, 100.0% similarity) APG08241 (95.22%identity, 96.93% similarity) APG07639 (94.88% identity, 96.93%similarity) US20130227743A1_100 (93.52% identity, 97.27% similarity)APG02279 (91.06% identity, 93.38% similarity) US_2016_0304898_A1-228(80.87% identity, 88.26% similarity) APG00006 (79.02% identity, 86.23%similarity) APG00107 (77.36% identity, 88.85% similarity) APG00749(76.74% identity, 85.38% similarity) APG00930 (76.35% identity, 87.16%similarity) APG00201 (75.74% identity, 84.59% similarity) APG00847(75.08% identity, 84.92% similarity) US20130227743A1_60 (41.58%identity, 46.86% similarity) APG08973 45, 50, 55, 60, 65, APG03440 70,75, 80, 85, 90, (93.31% identity, 95, 96, 97, 98, 99 93.65% similarity)US_2016_0311864_A1-91 (30.19% identity, 41.68% similarity)WP_017762581.1 (29.07% identity, 43.32% similarity) WP_044306759.1(29.07% identity, 42.17% similarity) AGP17992.1 (29.07% identity, 42.02%similarity) APG08990 95, 96, 97, 98, 99 APG06989 (96.79% identity,97.76% similarity) APG02921 (96.47% identity, 97.44% similarity)APG05969 (90.06% identity, 94.87% similarity) APG04226 (89.1% identity,94.55% similarity) WP_025150761.1 (88.14% identity, 93.27% similarity)SFS68380.1 (84.71% identity, 90.45% similarity) WP_016131662.1 (82.37%identity, 89.74% similarity) Cry55Aa1 (18.23% identity, 33.25%similarity) APG09256 98, 99 WP_000586614.1 (95.67% identity, 97.24%similarity) WP_065230018.1 (94.49% identity, 96.46% similarity)WP_000586616.1 (94.09% identity, 96.46% similarity) APG01269 (93.7%identity, 96.06% similarity) APG02245 (93.7% identity, 96.06%similarity) APG01103 (92.91% identity, 96.46% similarity) APG09842(92.52% identity, 96.46% similarity) Cry46Ab (30.0% identity, 44.19%similarity) APG09842 100 WP_000586617.1 (97.23% identity, 99.21%similarity) WP_044585299.1 (95.65% identity, 98.81% similarity) APG02429(95.65% identity, 98.02% similarity) WP_000586615.1 (95.65% identity,98.02% similarity) APG01103 (94.86% identity, 98.42% similarity)APG00737 (93.28% identity, 96.44% similarity) APG02768 (92.49% identity,98.02% similarity) Cry46Ab (29.64% identity, 44.3% similarity) APG0296095, 96, 97, 98, 99 APG04643 (97.84% identity, 99.38% similarity)WP_044444098.1 (92.28% identity, 92.90% similarity) APG00743 (88.89%identity, 91.36% similarity) APG02555 (87.65% identity, 91.05%similarity) APG08085 (84.26% identity, 89.51% similarity) WP_060749709.1(81.85% identity, 87.08% similarity) APG01265.0 98, 99, 100 APG04483.0(98.03% identity, 99.34% similarity) APG06465.0 (97.38% identity, 99.02%similarity) APG00696.0 US_2016_0355842_A1-160 (95.08% identity, 97.38%similarity) APG01882.0 (94.77% identity, 97.06% similarity) APG08138.0(94.1% identity, 97.38% similarity) APG03715.0 (90.62% identity, 93.44%similarity) APG04598.0 (78.69% identity, 92.13% similarity) APG00769.0(47.21% identity, 63.93% similarity) APG00535.0 US_2016_0355842_A1-118(46.25% identity, 62.54% similarity) APG00016.0 US_2016_0304898_A1-26(44.27% identity, 59.13% similarity) APG03954.0 99, 100 APG00426.0US_2016_0355842_A1-88 (98.12% identity, 98.93% similarity) APG01022.0(98.12% identity, 98.12% similarity) APG08225.0 (98.12% identity, 98.12%similarity) APG02400.0 (97.86% identity, 98.12% similarity) APG00891.0US_2016_0355842_A1-184 (92.58% identity, 93.61% similarity) APG00227.0US_2016_0311864_A1-83 (90.77% identity, 91.27% similarity) APG00517.0US_2016_0311864_A1-187 (81.02% identity, 82.89% similarity) APG00158.0US_2016_0355842_A1-8 (63.78% identity, 71.92% similarity) APG01231.0(57.44% identity, 67.36% similarity) WP_071770709.1 (57.33% identity,67.54% similarity) APG00253.0 US_2016_0355842_A1-39 (57.11% identity,68.42% similarity) APG00704.0 US_2016_0355842_A1-161 (56.92% identity,67.1% similarity) APG00020.0 US_2016_0304898_A1-33 (56.05% identity,68.42% similarity) APG01508.0 (56.05% identity, 68.42% similarity)APG00448.0 US_2016_0366881_A1-70 (56.05% identity, 66.58% similarity)APG00418.0 US_2016_0355842_A1-84 (55.76% identity, 66.75% similarity)APG00481.0 US_2016_0355842_A1-103 (55.14% identity, 65.41% similarity)APG00764.0 US_2016_0355842_A1-168 (55.03% identity, 66.08% similarity)APG04375.0 92, 93, 94, 95, 96, APG04793.0 97, 98, 99, 100 (91.14%identity, 94.1% similarity) APG03686.0 (89.67% identity, 94.1%similarity) APG07676.0 (88.93% identity, 94.1% similarity) APG03747.0(86.72% identity, 92.99% similarity) APG08525.0 (84.13% identity, 91.51%similarity) APG00014.0 US_2016_0304898_A1-24 (50.18% identity, 66.3%similarity) APG04013.0 (48.6% identity, 65.03% similarity)WP_033694890.1 (34.05% identity, 45.41% similarity) US_9328356_B2-22(32.98% identity, 48.58% similarity) APG00614.0 US_2016_0355842_A1-144(32.62% identity, 49.29% similarity) AAL26870.1 (31.14% identity, 46.37%similarity) APG02740.0 (30.3% identity, 51.18% similarity) APG04900.050, 55, 60, 65, 70, APG02225.0 75, 80, 85, 90, 91, (95.34% identity, 92,93, 94, 95, 96, 97.25% similarity) 97, 98, 99, 100 APG00156.0US_2016_0355842_A1-6 (33.14% identity, 46.08% similarity)US_8318900_B2-91 (29.66% identity, 44.23% similarity) APG04067.0 (25.81%identity, 36.59% similarity) APG00770.0 96, 97, 98, 99, 100US_2016_0339078_A1-28662 (94.61% identity, 95.59% similarity)WP_018782198.1 (94.61% identity, 95.59% similarity) WP_033798332.1(94.61% identity, 95.59% similarity) WP_040119538.1 (94.61% identity,95.59% similarity) WP_016114769.1 (94.61% identity, 95.1% similarity)WP_018766353.1 (94.61% identity, 95.1% similarity) APG05707.0 (94.12%identity, 95.1% similarity) US_2016_0339078_A1-28716 (94.12% identity,95.1% similarity) APG09444.0 (93.63% identity, 96.57% similarity)APG01987.0 (93.63% identity, 94.61% similarity) SDY81229.1 (92.16%identity, 94.12% similarity) APG01989.0 (91.67% identity, 93.63%similarity) WP_026594351.1 (91.67% identity, 93.63% similarity)APG00456.0 US_2016_0311864_A1-180 (91.18% identity, 92.65% similarity)APG01577.0 98, 99, 100 WP_043938562.1 (98.81% identity, 100.0%similarity) APG02248.0 (97.23% identity, 100.0% similarity)WP_016082893.1 (97.23% identity, 100.0% similarity) WP_048564006.1(95.65% identity, 98.42% similarity) WP_061667036.1 (95.26% identity,98.42% similarity) WP_000586615.1 (94.86% identity, 98.02% similarity)OUA56120.1 (94.86% identity, 96.84% similarity) ADQ73630.1 (94.47%identity, 98.02% similarity) WP_000586614.1 (94.47% identity, 98.02%similarity) WP_044585299.1 (94.47% identity, 98.02% similarity)OUC00742.1 (94.47% identity, 97.63% similarity) APG00737.0 (94.47%identity, 96.44% similarity) APG02245.0 (94.07% identity, 97.63%similarity) APG05399.0 (94.07% identity, 97.63% similarity)US_9403881_B2-6 (94.07% identity, 97.63% similarity) APG09892.0 (93.7%identity, 96.85% similarity) APG02768.0 (93.68% identity, 98.02%similarity) APG09446.0 (93.68% identity, 97.23% similarity) APG09857.0(93.68% identity, 97.23% similarity) APG09842.0 (93.28% identity, 97.63%similarity) APG01269.0 (92.91% identity, 96.06% similarity) APG01103.0(92.89% identity, 98.02% similarity) APG02429.0 (92.89% identity, 97.23%similarity) APG02756.0 (92.89% identity, 96.84% similarity) APG03148.0(92.49% identity, 96.44% similarity) APG09256.0 (91.73% identity, 96.06%similarity) APG01797.0 5, 10, 15, 20, 25, 30, APG03217.0 35, 40, 45, 50,55, (96.9% identity, 60, 65, 70, 75, 80, 98.19% similarity) 85, 90, 91,92, 93, 94, 95, 96, 97, 98, 99, 100 APG08029.0 (96.9% identity, 97.16%similarity) OUB22522.1 (30.18% identity, 30.45% similarity) APG07676.0(25.19% identity, 38.42% similarity) OUB46342.1 (25.06% identity, 39.9%similarity) APG01987.0 100 US_2016_0339078_A1-28662 (97.55% identity,99.02% similarity) WP_033798332.1 (97.55% identity, 99.02% similarity)WP_040119538.1 (97.55% identity, 99.02% similarity) WP_016114769.1(97.55% identity, 98.53% similarity) WP_018766353.1 (97.55% identity,98.53% similarity) WP_018782198.1 (97.06% identity, 99.02% similarity)US_2016_0339078_A1-28716 (97.06% identity, 98.53% similarity) APG05707.0(96.08% identity, 97.06% similarity) APG09444.0 (94.61% identity, 98.04%similarity) SDY81229.1 (93.63% identity, 96.08% similarity) APG00770.0(93.63% identity, 94.61% similarity) APG00456.0 US_2016_0311864_A1-180(93.14% identity, 95.59% similarity) APG01989.0 (93.14% identity, 95.59%similarity) WP_026594351.1 (93.14% identity, 95.59% similarity)APG09717.0 (91.67% identity, 94.61% similarity) APG02248.0 98, 99, 100WP_016082893.1 (100.0% identity, 100.0% similarity) WP_043938562.1(98.42% identity, 100.0% similarity) APG01577.0 (97.23% identity, 100.0%similarity) WP_061667036.1 (95.65% identity, 98.42% similarity)WP_000586614.1 (95.65% identity, 98.02% similarity) APG05399.0 (95.26%identity, 97.63% similarity) OTW75879.1 (95.26% identity, 97.63%similarity) OUB65258.1 (95.26% identity, 97.63% similarity)WP_065230018.1 (95.26% identity, 97.23% similarity) ADQ73630.1 (94.86%identity, 98.02% similarity) OTW93744.1 (94.86% identity, 97.63%similarity) APG09857.0 (94.86% identity, 97.23% similarity) APG02245.0(94.47% identity, 97.63% similarity) APG09892.0 (94.09% identity, 96.85%similarity) APG09446.0 (94.07% identity, 97.23% similarity) OUA56120.1(93.68% identity, 96.84% similarity) APG01103.0 (93.28% identity, 98.02%similarity) APG02768.0 (93.28% identity, 97.63% similarity) APG00737.0(93.28% identity, 96.44% similarity) APG01269.0 (92.91% identity, 96.06%similarity) APG09256.0 (92.91% identity, 96.06% similarity)US_9403881_B2-6 (92.89% identity, 97.63% similarity) APG09842.0 (92.49%identity, 97.23% similarity) APG02429.0 (92.09% identity, 96.84%similarity) APG02756.0 (92.09% identity, 96.44% similarity) APG03148.0(92.09% identity, 96.05% similarity) APG02443.0 94, 95, 96, 97, 98,WP_078205335.1 99, 100 (99.6% identity, 99.6% similarity) SCA96918.1(98.02% identity, 98.02% similarity) APG01150.0 (97.63% identity, 98.02%similarity) WP_061530406.1 (97.63% identity, 98.02% similarity)WP_064474064.1 (97.63% identity, 98.02% similarity) APG03040.0 (97.23%identity, 98.02% similarity) APG03017.0 (97.23% identity, 97.63%similarity) APG00788.0 (96.84% identity, 97.63% similarity) APG05678.0(96.84% identity, 97.63% similarity) WP_016096697.1 (96.84% identity,97.23% similarity) APG02756.0 98, 99, 100 APG03148.0 (98.42% identity,99.6% similarity) WP_000586617.1 (98.02% identity, 99.21% similarity)APG09842.0 (97.63% identity, 99.21% similarity) WP_044585299.1 (96.44%identity, 98.81% similarity) APG02429.0 (95.65% identity, 98.02%similarity) WP_000586615.1 (95.65% identity, 98.02% similarity)WP_048564006.1 (94.86% identity, 98.42% similarity) WP_078994697.1(94.86% identity, 97.23% similarity) WP_065230018.1 (94.47% identity,98.42% similarity) WP_000586616.1 (94.47% identity, 96.84% similarity)OUC00742.1 (94.07% identity, 97.63% similarity) US_9403881_B2-6 (94.07%identity, 97.63% similarity) WP_016090629.1 (94.07% identity, 97.23%similarity) APG01103.0 (93.68% identity, 97.63% similarity) APG02768.0(93.28% identity, 98.02% similarity) APG09857.0 (93.28% identity, 97.63%similarity) APG01577.0 (92.89% identity, 96.84% similarity) APG00737.0(92.89% identity, 95.65% similarity) APG09256.0 (92.13% identity, 95.67%similarity) APG02248.0 (92.09% identity, 96.44% similarity) APG05399.0(92.09% identity, 96.44% similarity) APG09892.0 (91.73% identity, 96.46%similarity) APG01269.0 (91.34% identity, 95.67% similarity) APG02245.0(91.3% identity, 96.44% similarity) APG09446.0 (90.91% identity, 96.05%similarity) APG03017.0 94, 95, 96, 97, 98, WP_016096697.1 99, 100 (99.6%identity, 99.6% similarity) APG02443.0 (97.23% identity, 97.63%similarity) SCA96918.1 (96.84% identity, 97.23% similarity)WP_078205335.1 (96.84% identity, 97.23% similarity) APG01150.0 (96.44%identity, 97.23% similarity) WP_061530406.1 (96.44% identity, 97.23%similarity) WP_064474064.1 (96.44% identity, 97.23% similarity)APG03040.0 (96.05% identity, 97.23% similarity) APG00788.0 (95.65%identity, 96.84% similarity) APG05678.0 (95.65% identity, 96.84%similarity) APG03148.0 98, 99, 100 APG02756.0 (98.42% identity, 99.6%similarity) WP_000586617.1 (98.02% identity, 98.81% similarity)APG09842.0 (96.84% identity, 98.81% similarity) WP_044585299.1 (96.44%identity, 98.42% similarity) APG02429.0 (95.65% identity, 97.63%similarity) WP_078994697.1 (95.65% identity, 96.84% similarity)WP_000586615.1 (95.26% identity, 97.63% similarity) WP_000586616.1(95.26% identity, 96.44% similarity) WP_065230018.1 (94.86% identity,98.02% similarity) WP_048564006.1 (94.47% identity, 98.02% similarity)APG02768.0 (94.07% identity, 97.63% similarity) APG01103.0 (94.07%identity, 97.23% similarity) OUC00742.1 (94.07% identity, 97.23%similarity) APG09857.0 (93.68% identity, 97.23% similarity)US_9403881_B2-6 (93.68% identity, 97.23% similarity) WP_016090629.1(93.68% identity, 96.84% similarity) APG00737.0 (92.89% identity, 95.26%similarity) APG09256.0 (92.52% identity, 95.28% similarity) APG01577.0(92.49% identity, 96.44% similarity) APG05399.0 (92.49% identity, 96.05%similarity) APG09892.0 (92.13% identity, 96.06% similarity) APG02248.0(92.09% identity, 96.05% similarity) APG01269.0 (91.73% identity, 95.28%similarity) APG02245.0 (91.7% identity, 96.05% similarity) APG09446.0(91.3% identity, 95.65% similarity) APG03574.0 60, 65, 70, 75, 80,APG04686.0 85, 90, 91, 92, 93, (98.72% identity, 94, 95, 96, 97, 98,99.57% similarity) 99, 100 APG03238.0 (98.5% identity, 99.14%similarity) APG08151.0 (98.5% identity, 99.14% similarity) APG06338.0(97.43% identity, 98.5% similarity) WP_082530295.1 (96.79% identity,97.43% similarity) APG03831.0 (95.93% identity, 97.86% similarity)WP_081932411.1 (95.72% identity, 96.79% similarity) WP_076565151.1(94.86% identity, 96.36% similarity) WP_071174111.1 (93.79% identity,95.93% similarity) WP_080769331.1 (93.58% identity, 95.5% similarity)KIF65774.1 (92.29% identity, 94.22% similarity) APG05399.0 97, 98, 99,100 WP_000586614.1 (97.23% identity, 98.02% similarity) OTW75879.1(96.84% identity, 97.63% similarity) OUB65258.1 (96.84% identity, 97.63%similarity) OTW93744.1 (96.44% identity, 97.63% similarity) APG09857.0(96.44% identity, 97.23% similarity) APG02245.0 (96.05% identity, 97.63%similarity) WP_065230018.1 (96.05% identity, 97.23% similarity)APG09446.0 (95.65% identity, 97.23% similarity) APG09256.0 (95.28%identity, 96.85% similarity) APG02248.0 (95.26% identity, 97.63%similarity) WP_016082893.1 (95.26% identity, 97.63% similarity)WP_043938562.1 (94.47% identity, 97.63% similarity) WP_061667036.1(94.47% identity, 97.63% similarity) WP_078994697.1 (94.47% identity,96.84% similarity) APG09892.0 (94.09% identity, 96.06% similarity)APG01577.0 (94.07% identity, 97.63% similarity) ADQ73630.1 (93.68%identity, 97.23% similarity) APG01269.0 (92.91% identity, 95.28%similarity) APG09842.0 (92.49% identity, 97.23% similarity) APG03148.0(92.49% identity, 96.05% similarity) APG01103.0 (92.09% identity, 97.23%similarity) APG02768.0 (92.09% identity, 96.84% similarity) APG02756.0(92.09% identity, 96.44% similarity) APG00737.0 (92.09% identity, 94.86%similarity) APG02429.0 (90.91% identity, 96.05% similarity)US_9403881_B2-6 (90.91% identity, 96.05% similarity) APG05678.0 93, 94,95, 96, 97, APG03040.0 98, 99, 100 (98.81% identity, 98.81% similarity)WP_064474064.1 (98.42% identity, 98.81% similarity) APG01150.0 (97.63%identity, 98.02% similarity) WP_061530406.1 (97.63% identity, 98.02%similarity) APG00788.0 (96.84% identity, 97.63% similarity) APG02443.0(96.84% identity, 97.63% similarity) SCA96918.1 (96.44% identity, 97.23%similarity) WP_078205335.1 (96.44% identity, 97.23% similarity)APG03017.0 (95.65% identity, 96.84% similarity) WP_016096697.1 (95.26%identity, 96.44% similarity) APG05707.0 99, 100 SDY81229.1 (96.57%identity, 98.04% similarity) US_2016_0339078_A1-28662 (96.57% identity,98.04% similarity) WP_033798332.1 (96.57% identity, 98.04% similarity)WP_040119538.1 (96.57% identity, 98.04% similarity) WP_016114769.1(96.57% identity, 97.55% similarity) WP_018766353.1 (96.57% identity,97.55% similarity) WP_018782198.1 (96.08% identity, 98.04% similarity)APG01989.0 (96.08% identity, 97.55% similarity) US_2016_0339078_A1-28716(96.08% identity, 97.55% similarity) WP_026594351.1 (96.08% identity,97.55% similarity) APG01987.0 (96.08% identity, 97.06% similarity)APG00456.0 US_2016_0311864_A1-180 (95.1% identity, 97.06% similarity)APG09717.0 (94.61% identity, 96.08% similarity) APG00770.0 (94.12%identity, 95.1% similarity) APG09444.0 (93.63% identity, 97.06%similarity) APG06385.0 95, 96, 97, 98, 99, APG02923.0 100 (96.67%identity, 96.67% similarity) WP_016110459.1 (94.58% identity, 95.83%similarity) APG00128.0 US_2016_0304898_A1-157 (90.83% identity, 94.58%similarity) APG00177.0 US_2016_0311864_A1-52 (55.6% identity, 69.11%similarity) APG00126.0 US_2016_0304898_A1-153 (54.15% identity, 69.17%similarity) APG00437.0 US_2016_0366881_A1-62 (53.23% identity, 67.68%similarity) APG00121.0 US_2016_0311864_A1-29 (52.31% identity, 62.28%similarity) WP_016110460.1 (51.97% identity, 67.72% similarity)APG01246.0 (42.86% identity, 59.85% similarity) APG00168.0US_2016_0311864_A1-44 (42.18% identity, 57.48% similarity) APG08372.065, 70, 75, 80, 85, WP_061688274.1 90, 91, 92, 93, 94, (100.0% identity,95, 96, 97, 98, 99, 100.0% similarity) 100 APG02674.0 (96.26% identity,97.13% similarity) APG01420.0 (93.1% identity, 95.69% similarity)WP_050595402.1 (90.52% identity, 91.38% similarity) WP_001267112.1(50.0% identity, 60.92% similarity) WP_023524027.1 (49.86% identity,65.1% similarity) US_8461415_B2-32 (49.3% identity, 60.28% similarity)WP_043924590.1 (47.31% identity, 60.75% similarity)US_2016_0017363_A1-59 (46.11% identity, 58.33% similarity)WP_078401252.1 (45.63% identity, 61.97% similarity) APG09444.0 99, 100APG01987.0 (94.61% identity, 98.04% similarity) US_2016_0339078_A1-28662(94.61% identity, 98.04% similarity) WP_033798332.1 (94.61% identity,98.04% similarity) WP_040119538.1 (94.61% identity, 98.04% similarity)WP_016114769.1 (94.61% identity, 97.55% similarity) WP_018766353.1(94.61% identity, 97.55% similarity) WP_018782198.1 (94.12% identity,98.04% similarity) US_2016_0339078_A1-28716 (94.12% identity, 97.55%similarity) APG05707.0 (93.63% identity, 97.06% similarity) APG00770.0(93.63% identity, 96.57% similarity) SDY81229.1 (91.67% identity, 96.08%similarity) APG00456.0 US_2016_0311864_A1-180 (91.67% identity, 95.59%similarity) APG01989.0 (91.18% identity, 95.59% similarity)WP_026594351.1 (91.18% identity, 95.59% similarity) APG09446.0 96, 97,98, 99, 100 APG02245.0 (99.6% identity, 99.6% similarity) WP_000586614.1(96.05% identity, 97.63% similarity) APG05399.0 (95.65% identity, 97.23%similarity) OTW75879.1 (95.65% identity, 97.23% similarity) OUB65258.1(95.65% identity, 97.23% similarity) OTW93744.1 (95.26% identity, 97.23%similarity) APG09857.0 (95.26% identity, 96.84% similarity)WP_065230018.1 (94.86% identity, 96.84% similarity) APG02248.0 (94.07%identity, 97.23% similarity) WP_016082893.1 (94.07% identity, 97.23%similarity) WP_043938562.1 (94.07% identity, 97.23% similarity)APG01577.0 (93.68% identity, 97.23% similarity) WP_061667036.1 (93.68%identity, 97.23% similarity) APG09256.0 (93.31% identity, 95.67%similarity) WP_078994697.1 (93.28% identity, 96.44% similarity)APG09892.0 (92.91% identity, 95.67% similarity) ADQ73630.1 (92.89%identity, 96.84% similarity) APG01269.0 (92.52% identity, 94.88%similarity) APG00737.0 (91.7% identity, 94.47% similarity) APG01103.0(91.3% identity, 96.84% similarity) APG09842.0 (91.3% identity, 96.84%similarity) APG02768.0 (91.3% identity, 96.44% similarity) APG03148.0(91.3% identity, 95.65% similarity) APG02756.0 (90.91% identity, 96.05%similarity) APG02429.0 (90.12% identity, 95.65% similarity) APG09857.098, 99, 100 WP_000586614.1 (98.42% identity, 99.21% similarity)WP_065230018.1 (98.02% identity, 99.21% similarity) OTW75879.1 (98.02%identity, 98.81% similarity) OTW93744.1 (97.63% identity, 98.81%similarity) OUB65258.1 (97.23% identity, 98.02% similarity) APG05399.0(96.44% identity, 97.23% similarity) APG02245.0 (95.65% identity, 97.23%similarity) APG09892.0 (95.28% identity, 97.24% similarity)WP_061667036.1 (95.26% identity, 98.02% similarity) APG09446.0 (95.26%identity, 96.84% similarity) APG09256.0 (94.88% identity, 96.46%similarity) APG02248.0 (94.86% identity, 97.23% similarity)WP_016082893.1 (94.86% identity, 97.23% similarity) WP_078994697.1(94.86% identity, 97.23% similarity) WP_000586617.1 (94.47% identity,98.42% similarity) ADQ73630.1 (94.47% identity, 97.63% similarity)APG01577.0 (93.68% identity, 97.23% similarity) APG03148.0 (93.68%identity, 97.23% similarity) APG01269.0 (93.31% identity, 95.67%similarity) APG02768.0 (93.28% identity, 98.02% similarity) APG02756.0(93.28% identity, 97.63% similarity) APG09842.0 (93.28% identity, 97.63%similarity) APG01103.0 (92.49% identity, 97.63% similarity) APG00737.0(92.49% identity, 95.26% similarity) APG02429.0 (92.09% identity, 97.23%similarity) US_9403881_B2-6 (92.09% identity, 97.23% similarity)APG09892.0 97, 98, 99, 100 WP_061667036.1 (96.06% identity, 97.64%similarity) APG01269.0 (95.67% identity, 96.85% similarity) ADQ73630.1(95.28% identity, 97.24% similarity) APG09857.0 (95.28% identity, 97.24%similarity) WP_000586614.1 (95.28% identity, 97.24% similarity)APG02768.0 (94.88% identity, 98.43% similarity) WP_065230018.1 (94.88%identity, 97.24% similarity) OTW75879.1 (94.88% identity, 96.85%similarity) OUB65258.1 (94.88% identity, 96.85% similarity) OTW93744.1(94.49% identity, 96.85% similarity) APG02248.0 (94.09% identity, 96.85%similarity) APG09256.0 (94.09% identity, 96.85% similarity)WP_016082893.1 (94.09% identity, 96.85% similarity) WP_043938562.1(94.09% identity, 96.85% similarity) APG05399.0 (94.09% identity, 96.06%similarity) WP_044585299.1 (93.7% identity, 97.64% similarity)APG01577.0 (93.7% identity, 96.85% similarity) APG02245.0 (93.31%identity, 96.06% similarity) APG09446.0 (92.91% identity, 95.67%similarity) APG01103.0 (92.52% identity, 97.24% similarity) APG00737.0(92.52% identity, 94.88% similarity) APG02429.0 (92.13% identity, 96.85%similarity) US_9403881_B2-6 (92.13% identity, 96.85% similarity)APG03148.0 (92.13% identity, 96.06% similarity) APG02756.0 (91.73%identity, 96.46% similarity) APG09842.0 (91.73% identity, 96.46%similarity) APG01172 40, 45, 50, 55, 60, APG00799 65, 70, 75, 80, 85,(84.88% identity, 90, 95, 96, 97, 98, 99 89.44% similarity) C0ZKJ5_BREBN(22.24% identity, 37.37% similarity) Cry5Adl (21.31% identity, 32.25%similarity) APG01288 45, 50, 55, 60, 65, APG00801 70, 75, 80, 85, 90,(57.05% identity, 95, 96, 97, 98, 99 66.46% similarity) APG00099 -US_2016_0177333_A1-55 (55.75% identity, 65.90% similarity) APG00784(55.74% identity, 65.45% similarity) Cry21Aa2 (28.67% identity, 41.63%similarity) APG05711 55, 60, 65, 70, 75, APG00182 80, 85, 90, 95, 96,(66.1% identity, 97, 98, 99 75.79% similarity) CUB55361.1 (35.55%identity, 50.68% similarity)

i. Classes of Pesticidal Proteins

The pesticidal proteins provided herein and the nucleotide sequencesencoding them are useful in methods for impacting pests. That is, thecompositions and methods of the invention find use in agriculture forcontrolling or killing pests, including pests of many crop plants. Thepesticidal proteins provided herein are toxin proteins from bacteria andexhibit activity against certain pests. The pesticidal proteins are fromseveral classes of toxins including Cry, Cyt, BIN, Mtx toxins. See, forexample, Table 1 for the specific protein classifications of the variousSEQ ID NOs provided herein. In addition, reference is made throughoutthis disclosure to Pfam database entries. The Pfam database is adatabase of protein families, each represented by multiple sequencealignments and a profile hidden Markov model. Finn et al. (2014) Nucl.Acid Res. Database Issue 42:D222-D230.

Bacillus thuringiensis (Bt) is a gram-positive bacterium that producesinsecticidal proteins as crystal inclusions during its sporulation phaseof growth. The proteinaceous inclusions of Bacillus thuringiensis (Bt)are called crystal proteins or δ-endotoxins (or Cry proteins), which aretoxic to members of the class Insecta and other invertebrates.Similarly, Cyt proteins are parasporal inclusion proteins from Bt thatexhibits hemolytic (Cytolitic) activity or has obvious sequencesimilarity to a known Cyt protein. These toxins are highly specific totheir target organism, are innocuous to humans, vertebrates, and plants.

The structure of the Cry toxins reveals five conserved amino acidblocks, concentrated mainly in the center of the domain or at thejunction between the domains. The Cry toxin consists of three domains,each with a specific function. Domain I is a seven α-helix bundle inwhich a central helix is completely surrounded by six outer helices.This domain is implicated in channel formation in the membrane. DomainII appears as a triangular column of three anti-parallel β-sheets, whichare similar to antigen-binding regions of immunoglobulins. Domain IIIcontains anti-parallel β-strands in a β sandwich form. The N-terminalpart of the toxin protein is responsible for its toxicity andspecificity and contains five conserved regions. The C-terminal part isusually highly conserved and probably responsible for crystal formation.See, for example, U.S. Pat. No. 8,878,007.

Strains of B. thuringiensis show a wide range of specificity againstdifferent insect orders (Lepidoptera, Diptera, Coleoptera, Hymenoptera,Homoptera, Phthiraptera or Mallophaga, and Acari) and otherinvertebrates (Nemathelminthes, Platyhelminthes, and Sarocomastebrates).The cry proteins have been classified into groups based on toxicity tovarious insect and invertebrate groups. Generally, Cry I demonstratestoxicity to lepidopterans, Cry II to lepidopterans and dipterans, CryIIIto coleopterans, Cry IV to dipterans, and Cry V and Cry VI to nematodes.New Cry proteins can be identified and assigned to a Cry group based onamino acid identity. See, for example, Bravo, A. (1997) J. of Bacteriol.179:2793-2801; Bravo et al. (2013) Microb. Biotechnol. 6:17-26, hereinincorporated by reference.

Over 750 different cry gene sequences have been classified into 73groups (Cry1-Cry73), with new members of this gene family continuing tobe discovered (Crickmore et al. (2014) www.btnomenclature.info/). Thecry gene family consists of several phylogentically non-related proteinfamilies that may have different modes of action: the family ofthree-domain Cry toxins, the family of mosquitocidal Cry toxins, thefamily of the binary-like toxins, and the Cyt family of toxins (Bravo etal., 2005). Some Bt strains produce additional insecticidal toxins, theVIP toxins. See, also, Cohen et al. (2011) J. Mol. Biol. 413:4-814;Crickmore et al. (2014) Bacillus thuringiensis toxin nomenclature, foundon the world wide web at lifesci.sussex.ac.uk/home/Neil_CrickmoreSt/;Crickmore et al. (1988) Microbiol. Mol. Biol. Rev. 62: 807-813; Gill etal. (1992) Ann. Rev. Entomol. 37: 807-636; Goldbert et al. (1997) Appl.Environ. Microbiol. 63:2716-2712; Knowles et al. (1992) Proc. R. Soc.Ser. B. 248: 1-7; Koni et al. (1994) Microbiology 140: 1869-1880; Lailaket al. (2013) Biochem. Biophys. Res. Commun. 435: 216-221; Lopez-Diaz etal. (2013) Environ. Microbiol. 15: 3030-3039; Perez et al. (2007) Cell.Microbiol. 9: 2931-2937; Promdonkoy et al. (2003) Biochem. J. 374:255-259; Rigden (2009) FEBS Lett. 583: 1555-1560; Schnepf et al. (1998)Microbiol. Mol. Biol. Rev. 62: 775-806; Soberon et al. (2013) Peptides41: 87-93; Thiery et al. (1998) J. Am. Mosq. Control Assoc. 14: 472-476;Thomas et al. (1983) FEBS Lett. 154: 362-368; Wirth et al. (1997) Proc.Natl. Acad. Sci. U.S.A. 94: 10536-10540; Wirth et al (2005) Appl.Environ. Microbiol. 71: 185-189; and, Zhang et al. (2006) Biosci.Biotechnol. Biochem. 70: 2199-2204; each of which is herein incorporatedby reference in their entirety.

Cyt designates a parasporal crystal inclusion protein from Bacillusthuringiensis with cytolytic activity, or a protein with sequencesimilarity to a known Cyt protein. (Crickmore et al. (1998) Microbiol.Mol. Biol. Rev. 62: 807-813). The gene is denoted by cyt. These proteinsare different in structure and activity from Cry proteins (Gill et al.(1992) Annu. Rev. Entomol. 37: 615-636). The Cyt toxins were firstdiscovered in B. thuringiensis subspecies israelensis (Goldberg et al.(1977) Mosq. News. 37: 355-358). There are 3 Cyt toxin familiesincluding 11 holotype toxins in the current nomenclature (Crickmore etal. (2014) Bacillus thuringiensis toxin nomenclature found on the worldwide web at lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/). The majorityof the B. thuringiensis isolates with cyt genes show activity againstdipteran insects (particularly mosquitoes and black flies), but thereare also cyt genes that have been described in B. thuringiensis strainstargeting lepidopteran or coleopteran insects (Guerchicoff et al. (1997)Appl. Environ. Microbiol. 63: 2716-2721).

The structure of Cyt2A, solved by X-ray crystallography, shows a singledomain where two outer layers of α-helix wrap around a mixed β-sheet.Further available crystal structures of Cyt toxins support a conservedα-β structural model with two α-helix hairpins flanking a β-sheet corecontaining seven to eight β-strands. (Cohen et al. (2011) J. Mol. Biol.413: 80 4-814) Mutagenic studies identified β-sheet residues as criticalfor toxicity, while mutations in the helical domains did not affecttoxicity (Adang et al.; Diversity of Bacillus thuringiensis CrystalToxins and Mechanism of Action. In: T. S. Dhadialla and S. S. Gill, eds,Advances in Insect Physiology, Vol. 47, Oxford: Academic Press, 2014,pp. 39-87.) The representative domain of the Cyt toxin is a δ-endotoxin,Bac_thur_toxin (Pfam PF01338).

There are multiple proposed models for the mode of action of Cyt toxins,and it is still an area of active investigation. Some Cyt proteins(Cyt1A) have been shown to require the presence of accessory proteinsfor crystallization. Cyt1A and Cyt2A protoxins are processed bydigestive proteases at the same sites in the N- and C-termini to astable toxin core. Cyt toxins then interact with non-saturated membranelipids, such as phosphatidylcholine, phosphatidylethanolamine, andsphingomyelin. For Cyt toxins, pore-formation and detergent-likemembrane disruption have been proposed as non-exclusive mechanisms; andit is generally accepted that both may occur depending on toxinconcentration, with lower concentrations favoring oligomeric pores andhigher concentrations leading to membrane breaks. (Butko (2003) Appl.Environ. Microbiol. 69: 2415-2422) In the pore-formation model, the Cyttoxin binds to the cell membrane, inducing the formation ofcation-selective channels in the membrane vesicles leading tocolloid-osmotic lysis of the cell. (Knowles et al. (1989) FEBS Lett.244: 259-262; Knowles et al. (1992) Proc. R. Soc. Ser. B. 248: 1-7 andPromdonkoy et al. (2003) Biochem. J. 374: 255-259). In the detergentmodel, there is a nonspecific aggregation of the toxin on the surface ofthe lipid bilayer leading to membrane disassembly and cell death. (Butko(2003) supra; Manceva et al. (2005) Biochem. 44: 589-597).

Multiple studies have shown synergistic activity between Cyt toxins andother B. thuringiensis toxins, particularly the Cry, Bin, and Mtxtoxins. This synergism has even been shown to overcome an insect'sresistance to the other toxin. (Wirth 1997, Wirth 2005, Thiery 1998,Zhang 2006) The Cyt synergistic effect for Cry toxins is proposed toinvolve Cyt1A binding to domain II of Cry toxins in solution or on themembrane plane to promote formation of a Cry toxin pre-pore oligomer.Formation of this oligomer is independent of the Cyt oligomerization,binding or insertion. (Lailak 2013, Perez 2007, Lopez-Diaz 2013).

A number of pesticidal proteins unrelated to the Cry proteins areproduced by some strains of B. thuringiensis and B. cereus duringvegetative growth (Estruch et al. (1996) Proc Natl Acad Sci USA93:5389-5394; Warren et al. (1994) WO 94/21795). These vegetativeinsecticidal proteins, or Vips, do not form parasporal crystal proteinsand are apparently secreted from the cell. The Vips are presentlyexcluded from the Cry protein nomenclature because they are notcrystal-forming proteins. The term VIP is a misnomer in the sense thatsome B. thuringiensis Cry proteins are also produced during vegetativegrowth as well as during the stationary and sporulation phases, mostnotably Cry3Aa. The location of the Vip genes in the B. thuringiensisgenome has been reported to reside on large plasmids that also encodecry genes (Mesrati et al. (2005) FEMS Microbiol. Lett. 244(2):353-8). Aweb-site for the nomenclature of Bt toxins can be found on the worldwide web at lifesci.sussex.ac.uk with the path“/home/Neil_Crickmore/Bt/” and at: “btnomenclature.info/”. See also,Schnepf et al. (1998) Microbiol. Mol. Biol. Rev. 62(3):775-806. Suchreferences are herein incorporated by reference.

To date four categories of Vips have been identified. Some Vip genesform binary two-component protein complexes; an “A” component is usuallythe “active” portion, and a “B” component is usually the “binding”portion. (Pfam pfam.xfam.org/family/PF03495). The Vip1 and Vip4 proteinsgenerally contain binary toxin B protein domains. Vip2 proteinsgenerally contain binary toxin A protein domains.

The Vip1 and Vip2 proteins are the two components of a binary toxin thatexhibits toxicity to coleopterans. Vip1Aa1 and Vip2Aa1 are very activeagainst corn rootworms, particularly Diabrotica virgifera and Diabroticalongicornis (Han et al. (1999) Nat. Struct. Biol. 6:932-936; Warren G W(1997) “Vegetative insecticidal proteins: novel proteins for control ofcorn pests” In: Carozzi N B, Koziel M (eds) Advances in insect control,the role of transgenic plants; Taylor & Francis Ltd, London, pp 109-21).The membrane-binding 95 kDa Vip1 multimer provides a pathway for the 52kDa vip2 ADP-ribosylase to enter the cytoplasm of target western cornrootworm cells (Warren (1997) supra). The NAD-dependentADP-ribosyltransferase Vip2 likely modifies monomeric actin at Arg177 toblock polymerization, leading to loss of the actin cytoskeleton andeventual cell death due to the rapid subunit ex-change within actinfilaments in vivo (Carlier M. F. (1990) Adv. Biophys. 26:51-73).

Like Cry toxins, activated Vip3A toxins are pore-forming proteinscapable of making stable ion channels in the membrane (Lee et al. (2003)Appl. Environ. Microbiol. 69:4648-4657). Vip3 proteins are activeagainst several major lepidopteran pests (Rang et al. (2005) Appl.Environ. Microbiol. 71(10):6276-6281; Bhalla et al. (2005) FEMSMicrobiol. Lett. 243:467-472; Estruch et al. (1998) WO 9844137; Estruchet al. (1996) Proc Natl Acad Sci USA 93:5389-5394; Selvapandiyan et al.(2001) Appl. Environ Microbiol. 67:5855-5858; Yu et al. (1997) Appl.Environ Microbiol. 63:532-536). Vip3A is active against Agrotis ipsilon,Spodoptera frugiperda, Spodoptera exigua, Heliothis virescens, andHelicoverpa zea (Warren et al. (1996) WO 96/10083; Estruch et al. (1996)Proc Natl Acad Sci USA 93:5389-5394). Like Cry toxins, Vip3A proteinsmust be activated by proteases prior to recognition at the surface ofthe midgut epithelium of specific membrane proteins different from thoserecognized by Cry toxins.

The MTX family of toxin proteins is characterized by the presence of aconserved domain, ETX_MTX2 (pfam 03318). Members of this family sharesequence homology with the mosquitocidal toxins Mtx2 and Mtx3 fromBacillus sphaericus, as well as with the epsilon toxin ETX fromClostridium perfringens (Cole et al. (2004) Nat. Struct. Mol. Biol. 11:797-8; Thanabalu et al. (1996) Gene 170:85-9). The MTX-like proteins arestructurally distinct from the three-domain Cry toxins, as they have anelongated and predominately β-sheet-based structure. However, similar tothe three-domain toxins, the MTX-like proteins are thought to form poresin the membranes of target cells (Adang et al. (2014) supra). Unlike thethree-domain Cry proteins, the MTX-like proteins are much smaller inlength, ranging from 267 amino acids (Cry23) to 340 amino acids(Cry15A).

To date, only 15 proteins belonging to the family of MTX-like toxinshave been assigned Cry names, making this a relatively small classcompared to the three-domain Cry family (Crickmore et al. (2014) supra;Adang et al. (2014) supra). The members of the MTX-like toxin familyinclude Cry15, Cry23, Cry33, Cry38, Cry45, Cry46, Cry51, Cry60A, Cry60B,and Cry64. This family exhibits a range of insecticidal activity,including activity against insect pests of the Lepidopteran andColeopteran orders. Some members of this family may form binarypartnerships with other proteins, which may or may not be required forinsecticidal activity.

Cry15 is a 34 kDA protein that was identified in Bacillus thuringiensisserovar thompsoni HD542; it occurs naturally in a crystal together withan unrelated protein of approximately 40 kDa. The gene encoding Cry15and its partner protein are arranged together in an operon. Cry15 alonehas been shown to have activity against lepidopteran insect pestsincluding Manduca sexta, Cydia pomonella, and Pieris rapae, with thepresence of the 40 kDA protein having been shown to increase activity ofCry15 only against C. pomonella (Brown K. and Whiteley H. (1992) J.Bacteriol. 174:549-557; Naimov et al. (2008) Appl. Environ. Microbiol.74:7145-7151). Further studies are needed to elucidate the function ofthe partner protein of Cry15. Similarly, Cry23 is a 29 kDA protein thathas been shown to have activity against the coleopteran pests Triboliumcastaneum and Popillia japonica together with its partner protein Cry37(Donovan et al. (2000), U.S. Pat. No. 6,063,756).

New members of the MTX-like family are continuing to be identified. AnETX_MTX toxin gene was recently identified in the genome of Bacillusthuringiensis serovar tolworthi strain Na205-3. This strain was found tobe toxic against the lepidpoteran pest Helicoverpa armigera, and it alsocontained homologs of Cry1, Cry11, Vip1, Vip2, and Vip3 (Palma et al.(2014) Genome Announc. 2(2): e00187-14. Published online Mar. 13, 2014at doi: 10.1128/genomeA.00187-14; PMCID: PMC3953196). Because theMTX-like proteins have a unique domain structure relative to thethree-domain Cry proteins, they are believed to possess a unique mode ofaction, thereby making them a valuable tool in insect control and thefight against insect resistance.

Bacterial cells produce large numbers of toxins with diverse specificityagainst host and non-host organisms. Large families of binary toxinshave been identified in numerous bacterial families, including toxinsthat have activity against insect pests. (Poopathi and Abidha (2010) J.Physiol. Path. 1(3): 22-38). Lysinibacillus sphaericus (Ls), formerlyBacillus sphaericus, (Ahmed et al. (2007) Int. J. Syst. Evol. Microbiol.57:1117-1125) is well-known as an insect biocontrol strain. Ls producesseveral insecticidal proteins, including the highly potent binarycomplex BinA/BinB. This binary complex forms a parasporal crystal in Lscells and has strong and specific activity against dipteran insects,specifically mosquitos. In some areas, insect resistance to existing Lsmosquitocidal strains has been reported. The discovery of new binarytoxins with different target specificity or the ability to overcomeinsect resistance is of significant interest.

The Ls binary insecticidal protein complex contains two majorpolypeptides, a 42 kDa polypeptide and a 51 kDa polypepdide, designatedBinA and BinB, respectively (Ahmed et al. (2007) supra). The twopolypeptides act synergistically to confer toxicity to their targets.Mode of action involves binding of the proteins to receptors in thelarval midgut. In some cases, the proteins are modified by proteasedigestion in the larval gut to produce activated forms. The BinBcomponent is thought to be involved in binding, while the BinA componentconfers toxicity (Nielsen-LeRoux et al. (2001) Appl. Environ. Microbiol.67(11):5049-5054). When cloned and expressed separately, the BinAcomponent is toxic to mosquito larvae, while the BinB component is not.However, co-administration of the proteins markedly increases toxicity(Nielsen-LeRoux et al. (2001) supra).

A small number of Bin protein homologs have been described frombacterial sources. Priest et al. (1997) Appl. Environ. Microbiol.63(4):1195-1198 describe a hybridization effort to identify new Lsstrains, although most of the genes they identified encoded proteinsidentical to the known BinA/BinB proteins. The BinA protein contains adefined conserved domain known as the Toxin 10 superfamily domain. Thistoxin domain was originally defined by its presence in BinA and BinB.The two proteins both have the domain, although the sequence similaritybetween BinA and BinB is limited in this region (<40%). The Cry49Aaprotein, which also has insecticidal activity, also has this domain(described below).

The Cry48Aa/Cry49Aa binary toxin of Ls has the ability to kill Culexquinquefasciatus mosquito larvae. These proteins are in a proteinstructural class that has some similarity to the Cry protein complex ofBacillus thuringiensis (Bt), a well-known insecticidal protein family.The Cry34/Cry35 binary toxin of Bt is also known to kill insects,including Western corn rootworm, a significant pest of corn. Cry34, ofwhich several variants have been identified, is a small (14 kDa)polypeptide, while Cry35 (also encoded by several variants) is a 44 kDapolypeptide. These proteins have some sequence homology with theBinA/BinB protein group and are thought to be evolutionarily related(Ellis et al. (2002) Appl. Environ. Microbiol. 68(3):1137-1145).

Phosphoinositide phospholipase C proteins (PI-PLC; alsophosphotidylinositol phospholipase C) are members of the broader groupof phospholipase C proteins. Many of these proteins play important rolesin signal transduction as part of normal cell physiology. Severalimportant bacterial toxins also contain domains with similarity to theseproteins (Titball, R. W. (1993) Microbiological Reviews. 57(2):347-366).Importantly, these proteins are implicated in signal amplificationduring intoxication of insect cells by Bt Cry proteins (Valaitis, A. P.(2008) Insect Biochemistry and Molecular Biology. 38: 611-618).

The PI-PLC toxin class occurs in Bacillus isolates, commonly seen inco-occurrence with homologs to other described toxin classes, such asBinary Toxins. This class of sequences has homology tophosphatidylinositol phosphodiesterases (also referred to asphosphatidylinositol-specific phospholipase C—PI-PLC). The crystalstructure and its active site were solved for B. cereus PI-PLC by Heinzet al (Heinz, et. al., (1995) The EMBO Journal. 14(16): 3855-3863). Theroles of the B. cereus PI-PLC active site amino acid residues incatalysis and substrate binding were investigated by Gassier et al usingsite-directed mutagenesis, kinetics, and crystal structure analysis(Gassier, et. al., (1997) Biochemistry. 36(42):12802-13).

These PI-PLC toxin proteins contain a PLC-like phosphodiesterase, TIMbeta/alpha-barrel domain (IPR017946) and/or a Phospholipase C,phosphatidylinositol-specific, X domain (IPR000909) (also referred to asthe PI-PLC X-box domain). We have also seen proteins with these domainsin combination with other typical Bacillus protein toxin domains. Thislist includes most commonly a lectin domain (IPR000772), a sugar-bindingdomain that can be present in one or more copies and is thought to bindcell membranes, as well as the Insecticidal crystal toxin (IPR008872)(also referred to as Toxin10 or P42), which is the defining domain ofthe Binary Toxin.

Previously, toxins of this PI-PLC class were defined in U.S. Pat. No.8,318,900 B2 SEQ ID NOs 30 (DNA) and 79 (amino acid), in U.S. PatentPublication No. 20110263488A1 SEQ ID NOs 8 (DNA) and 9 (amino acid), andin U.S. Pat. No. 8,461,421B2 SEQ ID NOs 3 (DNA) and 63 (amino acid).

Provided herein are pesticidal proteins from these classes of toxins.The pesticidal proteins are classified by their structure, homology toknown toxins and/or their pesticidal specificity.

ii. Variants and Fragments of Pesticidal Proteins and PolynucleotidesEncoding the Same

Pesticidal proteins or polypeptides of the invention include those setforth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51,52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104,105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118,119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132,133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146,147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160,161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174,175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188,189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202,203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216,217, 218, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229,230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243,244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257,258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271,272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285,286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299,300, 301, 302, 303, 304, 305, 306, 307, 308, and/or 309 and fragmentsand variants thereof. By “pesticidal toxin” or “pesticidal protein” or“pesticidal polypeptide” is intended a toxin or protein or polypeptidethat has activity against one or more pests, including, insects, fungi,nematodes, and the like such that the pest is killed or controlled.

An “isolated” or “purified” polypeptide or protein, or biologicallyactive portion thereof, is substantially or essentially free fromcomponents that normally accompany or interact with the polypeptide orprotein as found in its naturally occurring environment. Thus, anisolated or purified polypeptide or protein is substantially free ofother cellular material, or culture medium when produced by recombinanttechniques, or substantially free of chemical precursors or otherchemicals when chemically synthesized. A protein that is substantiallyfree of cellular material includes preparations of protein having lessthan about 30%, 20%, 10%, 5%, or 1% (by dry weight) of contaminatingprotein. When the protein of the invention or biologically activeportion thereof is recombinantly produced, optimally culture mediumrepresents less than about 30%, 20%, 10%, 5%, or 1% (by dry weight) ofchemical precursors or non-protein-of-interest chemicals.

The term “fragment” refers to a portion of a polypeptide sequence of theinvention. “Fragments” or “biologically active portions” includepolypeptides comprising a sufficient number of contiguous amino acidresidues to retain the biological activity, i.e., have pesticidalactivity. Fragments of the pesticidal proteins include those that areshorter than the full-length sequences, either due to the use of analternate downstream start site, or due to processing that produces ashorter protein having pesticidal activity. Processing may occur in theorganism the protein is expressed in, or in the pest after ingestion ofthe protein. Examples of fragments of the proteins can be found inTable 1. A biologically active portion of a pesticidal protein can be apolypeptide that is, for example, 10, 25, 50, 100, 150, 200, 250 or moreamino acids in length of any one of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98,99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223,224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279,280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293,294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307,308, and/or 309. Such biologically active portions can be prepared byrecombinant techniques and evaluated for pesticidal activity. As usedhere, a fragment comprises at least 8 contiguous amino acids of SEQ IDNOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55,56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107,108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121,122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135,136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149,150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163,164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177,178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191,192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205,206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 218,219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232,233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246,247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260,261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274,275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288,289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302,303, 304, 305, 306, 307, 308, and/or 309.

Bacterial genes, including those encoding the pesticidal proteinsdisclosed herein, quite often possess multiple methionine initiationcodons in proximity to the start of the open reading frame. Often,translation initiation at one or more of these start codons will lead togeneration of a functional protein. These start codons can include ATGcodons. However, bacteria such as Bacillus sp. also recognize the codonGTG as a start codon, and proteins that initiate translation at GTGcodons contain a methionine at the first amino acid. On rare occasions,translation in bacterial systems can initiate at a TTG codon, though inthis event the TTG encodes a methionine. Furthermore, it is not oftendetermined a priori which of these codons are used naturally in thebacterium. Thus, it is understood that use of one of the alternatemethionine codons may also lead to generation of pesticidal proteins.These pesticidal proteins are encompassed in the present invention andmay be used in the methods disclosed herein. It will be understood that,when expressed in plants, it will be necessary to alter the alternatestart codon to ATG for proper translation.

In various embodiments the pesticidal proteins provided herein includeamino acid sequences deduced from the full-length nucleotide sequencesand amino acid sequences that are shorter than the full-length sequencesdue to the use of an alternate downstream start site. Thus, thenucleotide sequence of the invention and/or vectors, host cells, andplants comprising the nucleotide sequence of the invention (and methodsof making and using the nucleotide sequence of the invention) maycomprise a nucleotide sequence encoding an alternate start site.

It is recognized that modifications may be made to the pesticidalpolypeptides provided herein creating variant proteins. Changes designedby man may be introduced through the application of site-directedmutagenesis techniques. Alternatively, native, as yet-unknown or as yetunidentified polynucleotides and/or polypeptides structurally and/orfunctionally-related to the sequences disclosed herein may also beidentified that fall within the scope of the present invention.Conservative amino acid substitutions may be made in nonconservedregions that do not alter the function of the pesticidal proteins.Alternatively, modifications may be made that improve the activity ofthe toxin. Modification of Cry toxins by domain III swapping hasresulted in some cases in hybrid toxins with improved toxicities againstcertain insect species. Thus, domain III swapping could be an effectivestrategy to improve toxicity of Cry toxins or to create novel hybridtoxins with toxicity against pests that show no susceptibility to theparental Cry toxins. Site-directed mutagenesis of domain II loopsequences may result in new toxins with increased insecticidal activity.Domain II loop regions are key binding regions of initial Cry toxinsthat are suitable targets for the mutagenesis and selection of Crytoxins with improved insecticidal properties. Domain I of the Cry toxinmay be modified to introduce protease cleavage sites to improve activityagainst certain pests. Strategies for shuffling the three differentdomains among large numbers of cry genes and high through outputbioassay screening methods may provide novel Cry toxins with improved ornovel toxicities.

As indicated, fragments and variants of the polypeptides disclosedherein will retain pesticidal activity. Pesticidal activity comprisesthe ability of the composition to achieve an observable effectdiminishing the occurrence or an activity of the target pest, includingfor example, bringing about death of at least one pest, or a noticeablereduction in pest growth, feeding, or normal physiological development.Such decreases in numbers, pest growth, feeding or normal developmentcan comprise any statistically significant decrease, including, forexample a decrease of about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,50%, 55%, 60%, 65%, 70%, 75%, 85%, 90%, 95% or greater. The pesticidalactivity against one or more of the various pests provided herein,including, for example, pesticidal activity against Coleoptera, Diptera,Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera,Nematodes, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera,Trichoptera, etc., or any other pest described herein. It is recognizedthat the pesticidal activity may be different or improved relative tothe activity of the native protein, or it may be unchanged, so long aspesticidal activity is retained. Methods for measuring pesticidalactivity are provide elsewhere herein. See also, Czapla and Lang (1990)J. Econ. Entomol. 83:2480-2485; Andrews et al. (1988) Biochem. J.252:199-206; Marrone et al. (1985) J. of Economic Entomology 78:290-293;and U.S. Pat. No. 5,743,477, all of which are herein incorporated byreference in their entirety.

By “variants” is intended polypeptides having an amino acid sequencethat is at least about 60%, about 65%, about 70%, about 75%, about 80%,about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%,about 98% or about 99% identical to the amino acid sequence of any ofSEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53,54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105,106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119,120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133,134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147,148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161,162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175,176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189,190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203,204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217,218, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230,231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244,245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258,259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272,273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286,287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300,301, 302, 303, 304, 305, 306, 307, 308, and/or 309 and retain pesticidalactivity. Note, Table 1 provides non-limiting examples of variantpolypeptides (and polynucleotide encoding the same) for each of SEQ IDNOS: 1-309. A biologically active variant of a pesticidal polypeptide ofthe invention may differ by as few as about 1-15 amino acid residues, asfew as about 1-10, such as about 6-10, as few as 5, as few as 4, as fewas 3, as few as 2, or as few as 1 amino acid residue. In specificembodiments, the polypeptides can comprise an N-terminal or a C-terminaltruncation, which can comprise at least a deletion of 10, 15, 20, 25,30, 35, 40, 45, 50 amino acids or more from either the N or C terminalof the polypeptide.

Table 2 provides protein domains found in SEQ ID NOs: 1-309 based onPFAM data. Both the domain description and the positions within a givenSEQ ID NO are provided in Table 2. In specific embodiments, the activevariant comprising any one of SEQ ID NOs: 1-309 can comprise at least70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequenceidentity to any one of SEQ ID NOs: 1-309 and further comprises at leastone of the conserved domain set forth in Table 2. For example, in oneembodiment, the active variant will comprise at least 70%, 75%, 76%,77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQID NO:1, and further comprises the native amino acids at positions18-252.

TABLE 2 Summary of PFAM domains in each of SEQ ID NOs: 1-309Modification PFAM Domain Domain positions APG ID Seq ID Type domainDescription Start Stop APG00589 Seq ID 1 PF03318 ETX MTX2 18 252APG00737 Seq ID 2 no PFAM domains APG00738 Seq ID 3 PF00388 PI-PLC-X 80200 APG00749 Seq ID 4 PF03318 ETX MTX2 29 254 APG00749 Seq ID 5Alternate start PF03318 ETX MTX2 27 252 modified (APG00749.1) APG00769Seq ID 6 PF03318 ETX MTX2 37 276 APG00788 Seq ID 7 no PFAM domainsAPG00790 Seq ID 8 PF01338 Bac thur toxin 22 247 APG00808 Seq ID 9 noPFAM domains APG00809 Seq ID 10 no PFAM domains APG00906 Seq ID 11 noPFAM domains APG00945 Seq ID 12 PF01338 Bac thur toxin 1 194 APG00955Seq ID 13 PF03318 ETX MTX2 26 255 APG00955 Seq ID 14 Alternate startPF03318 ETX MTX2 23 252 modified (APG00955.1) APG00965 Seq ID 15 PF03945Endotoxin N 78 291 PF00555 Endotoxin M 299 518 PF03944 Endotoxin C 528679 APG00974 Seq ID 16 PF03945 Endotoxin N 70 299 PF00555 Endotoxin M307 522 PF03944 Endotoxin C 532 672 APG00974 Seq ID 17 3′ TruncationPF03945 Endotoxin N 69 299 modified PF00555 Endotoxin M 307 522(APG00974.1) PF03944 Endotoxin C 532 671 APG00989 Seq ID 18 PF03318 ETXMTX2 67 324 APG00989 Seq ID 19 Signal PF03318 ETX MTX2 31 293 modifiedpeptide (APG00989.1) removed APG01022 Seq ID 20 PF03318 ETX MTX2 119 352APG01068 Seq ID 21 no PFAM domains APG01068 Seq ID 22 Signal no PFAMmodified peptide domains (APG01068.1) removed APG01078 Seq ID 23 PF03318ETX MTX2 9 119 APG01084 Seq ID 24 PF03945 Endotoxin N 77 278 APG01103Seq ID 25 no PFAM domains APG01121 Seq ID 26 PF01338 Bac thur toxin 17240 APG01150 Seq ID 27 no PFAM domains APG01199 Seq ID 28 PF03945Endotoxin N 60 251 PF00555 Endotoxin M 313 510 PF03944 Endotoxin C 520653 APG01199 Seq ID 29 3′ Truncation PF03945 Endotoxin N 60 251 modifiedPF00555 Endotoxin M 313 510 (APG01199.1) PF03944 Endotoxin C 520 652APG01242 Seq ID 30 no PFAM domains APG01242 Seq ID 31 Alternate start noPFAM modified domains (APG01242.1) APG01246 Seq ID 32 PF01338 Bac thurtoxin 27 240 APG01246 Seq ID 33 Alternate start PF01338 Bac thur toxin19 232 modified (APG01246.1) APG01257 Seq ID 34 PF01338 Bac thur toxin23 247 APG01269 Seq ID 35 no PFAM domains APG01273 Seq ID 36 no PFAMdomains APG01301 Seq ID 37 PF01338 Bac thur toxin 22 244 APG01401 Seq ID38 no PFAM domains APG01420 Seq ID 39 no PFAM domains APG01420 Seq ID 40Signal no PFAM modified peptide domains (APG01420.1) removed APG01451Seq ID 41 PF03318 ETX MTX2 1 198 APG01463 Seq ID 42 PF03318 ETX MTX2 86300 APG01507 Seq ID 43 no PFAM domains APG01507 Seq ID 44 Signal no PFAMmodified peptide domains (APG01507.1) removed APG01508 Seq ID 45 PF03318ETX MTX2 105 251 APG01508 Seq ID 46 Signal PF03318 ETX MTX2 55 222modified peptide (APG01508.1) removed APG01536 Seq ID 47 no PFAM domainsAPG01536 Seq ID 48 Signal no PFAM modified peptide domains (APG01536.1)removed APG01536 Seq ID 49 Alternate start no PFAM modified domains(APG01536.2) APG01700 Seq ID 50 no PFAM domains APG01700 Seq ID 51Alternate start no PFAM modified domains (APG01700.1) APG01882 Seq ID 52PF03318 ETX MTX2 40 281 APG01882 Seq ID 53 Alternate start PF03318 ETXMTX2 39 280 modified (APG01882.1) APG01944 Seq ID 54 PF03945 Endotoxin N417 588 APG01944 Seq ID 55 Alternate start PF03945 Endotoxin N 408 579modified (APG01944.1) APG01992 Seq ID 56 PF03945 Endotoxin N 78 295PF00555 Endotoxin M 309 521 PF03944 Endotoxin C 531 662 APG01992 Seq ID57 Alternate start PF03945 Endotoxin N 75 292 modified PF00555 EndotoxinM 306 518 (APG01992.1) PF03944 Endotoxin C 528 659 APG02038 Seq ID 58 noPFAM domains APG02067 Seq ID 59 PF03945 Endotoxin N 104 316 PF03944Endotoxin C 553 690 APG02067 Seq ID 60 Alternate start PF03945 EndotoxinN 77 289 modified PF03944 Endotoxin C 526 663 (APG02067.1) APG02067 SeqID 61 Alternate start PF03945 Endotoxin N 77 289 modified and 3′ PF00555Endotoxin M 297 516 (APG02067.2) Truncation PF03944 Endotoxin C 526 662APG02224 Seq ID 62 PF03945 Endotoxin N 76 288 PF00555 Endotoxin M 296504 PF03944 Endotoxin C 514 662 APG02224 Seq ID 63 3′ Truncation PF03945Endotoxin N 76 288 modified PF00555 Endotoxin M 296 504 (APG02224.1)PF03944 Endotoxin C 514 661 APG02225 Seq ID 64 PF01473 CW binding 1 362376 PF01473 CW binding 1 389 403 PF01473 CW binding 1 416 431 APG02280Seq ID 65 PF03318 ETX MTX2 86 323 APG02387 Seq ID 66 PF03945 Endotoxin N131 350 PF03944 Endotoxin C 539 677 APG02387 Seq ID 67 Signal PF03945Endotoxin N 99 318 modified peptide PF03944 Endotoxin C 507 645(APG02387.2) removed APG02387 Seq ID 68 Alternate start PF03945Endotoxin N 131 350 modified PF03944 Endotoxin C 539 677 (APG02387.1)APG02400 Seq ID 69 PF03318 ETX MTX2 119 352 APG02400 Seq ID 70 SignalPF03318 ETX MTX2 90 323 modified peptide (APG02400.1) removed APG02518Seq ID 71 no PFAM domains APG02518 Seq ID 72 Signal no PFAM modifiedpeptide domains (APG02518.1) removed APG02531 Seq ID 73 no PFAM domainsAPG02531 Seq ID 74 Alternate start no PFAM modified domains (APG02531.1)APG02552 Seq ID 75 PF03945 Endotoxin N 65 272 APG02552 Seq ID 76Alternate start PF03945 Endotoxin N 12 219 modified (APG02552.1)APG02555 Seq ID 77 no PFAM domains APG02555 Seq ID 78 Signal no PFAMmodified peptide domains (APG02555.1) removed APG02557 Seq ID 79 PF03318ETX MTX2 98 338 APG02557 Seq ID 80 Signal PF03318 ETX MTX2 64 305modified peptide (APG02557.1) removed APG02633 Seq ID 81 no PFAM domainsAPG02643 Seq ID 82 no PFAM domains APG02643 Seq ID 83 Alternate start noPFAM modified domains (APG02643.1) APG02740 Seq ID 84 no PFAM domainsAPG02740 Seq ID 85 Alternate start no PFAM modified domains (APG02740.1)APG02740 Seq ID 86 PF07029 CryBP1 18 169 CryBP1 (APG07203) APG02921 SeqID 87 no PFAM domains APG02921 Seq ID 88 Signal no PFAM modified peptidedomains (APG02921.1) removed APG02923 Seq ID 89 PF01338 Bac thur toxin15 234 APG02923 Seq ID 90 no PFAM domains APG03114 Seq ID 91 no PFAMdomains APG03114 Seq ID 92 Signal no PFAM modified peptide domains(APG03114.1) removed APG03238 Seq ID 93 PF06101 DUF946 19 271 APG03238Seq ID 94 Alternate start PF06101 DUF946 11 261 modified (APG03238.1)APG03440 Seq ID 95 PF03945 Endotoxin N 128 328 PF01473 CW binding 1 455470 PF01473 CW binding 1 487 506 APG03484 Seq ID 96 PF03318 ETX MTX2 158406 APG03484 Seq ID 97 Alternate start PF03318 ETX MTX2 119 368 modified(APG03484.1) APG03484 Seq ID 98 Signal PF03318 ETX MTX2 92 329 modifiedpeptide (APG03484.2) removed APG03619 Seq ID 99 PF03318 ETX MTX2 1 148APG03619 Seq ID Alternate start PF03318 ETX MTX2 1 148 modified 100(APG03619.1) APG03686 Seq ID no PFAM 101 domains APG03686 Seq IDAlternate start no PFAM modified 102 domains (APG03686.1) APG03715 SeqID PF03318 ETX MTX2 54 295 103 APG03715 Seq ID Alternate start PF03318ETX MTX2 39 296 modified 104 (APG03715.1) APG03747 Seq ID no PFAM 105domains APG03747 Seq ID Alternate start no PFAM modified 106 domains(APG03747.1) APG03760 Seq ID PF03318 ETX MTX2 63 327 107 APG03831 Seq IDPF06101 DUF946 14 273 108 APG03831 Seq ID Alternate start PF06101 DUF94611 263 modified 109 (APG03831.1) APG03867 Seq ID PF03318 ETX MTX2 38 265110 APG03867 Seq ID Alternate start PF03318 ETX MTX2 38 265 modified 111(APG03867.1) APG04067 Seq ID no PFAM 112 domains APG04067 Seq ID Signalno PFAM modified 113 peptide domains (APG04067.1) removed APG04099 SeqID PF03318 ETX MTX2 56 314 114 APG04099 Seq ID Signal PF03318 ETX MTX229 287 modified 115 peptide (APG04099.1) removed APG04152 Seq ID PF03945Endotoxin N 137 334 116 APG04152 Seq ID Alternate start PF03945Endotoxin N 120 317 modified 117 (APG04152.1) APG04450 Seq ID no PFAM118 domains APG04483 Seq ID PF03318 ETX MTX2 39 280 119 APG04686 Seq IDPF06101 DUF946 19 271 120 APG04686 Seq ID Alternate start PF06101 DUF94611 261 modified 121 (APG04686.1) APG04721 Seq ID no PFAM 122 domainsAPG04778 Seq ID PF03318 ETX MTX2 1 100 123 APG04793 Seq ID no PFAM 124domains APG04793 Seq ID Alternate start no PFAM modified 125 domains(APG04793.1) APG04925 Seq ID no PFAM 126 domains APG05213 Seq ID PF06101DUF946 12 223 127 APG05372 Seq ID no PFAM 128 domains APG05372 Seq IDAlternate start no PFAM modified 129 domains (APG05372.1) APG05500 SeqID PF03318 ETX MTX2 12 266 130 APG05500 Seq ID Alternate start PF03318ETX MTX2 12 266 modified 131 (APG05500.1) APG05553 Seq ID PF14200 RicinBlectin 2 2 84 132 APG05634 Seq ID PF03318 ETX MTX2 57 329 133 APG05706Seq ID PF03945 Endotoxin N 147 371 134 APG05706 Seq ID Alternate startPF03945 Endotoxin N 132 356 modified 135 (APG05706.1) APG06001 Seq ID noPFAM 136 domains APG06001 Seq ID Signal no PFAM modified 137 peptidedomains (APG06001.1) removed APG06281 Seq ID PF03318 ETX MTX2 18 111 138APG06324 Seq ID PF03318 ETX MTX2 69 330 139 APG06324 Seq ID SignalPF03318 ETX MTX2 34 290 modified 140 peptide (APG06324.1) removedAPG06338 Seq ID PF06101 DUF946 19 279 141 APG06338 Seq ID Alternatestart PF06101 DUF946 11 269 modified 142 (APG06338.1) APG06381 Seq ID noPFAM 143 domains APG06381 Seq ID Signal PF03318 ETX MTX2 9 221 modified144 peptide (APG06381.2) removed APG06381 Seq ID Alternate start no PFAMmodified 145 domains (APG06381.1) APG06465 Seq ID PF03318 ETX MTX2 39280 146 APG06501 Seq ID no PFAM 147 domains APG06501 Seq ID Signal noPFAM modified 148 peptide domains (APG06501.1) removed APG06589 Seq IDno PFAM 149 domains APG06589 Seq ID Alternate start no PFAM modified 150domains (APG06589.1) APG06676 Seq ID PF03318 ETX MTX2 58 266 151APG06894 Seq ID PF01338 Bac thur toxin 2 190 152 APG06894 Seq IDAlternate start PF01338 Bac thur toxin 1 190 modified 153 (APG06894.1)APG06989 Seq ID no PFAM 154 domains APG06989 Seq ID Signal no PFAMmodified 155 peptide domains (APG06989.2) removed APG06989 Seq IDAlternate start no PFAM modified 156 domains (APG06989.1) APG06997 SeqID PF03945 Endotoxin N 112 315 157 PF01473 CW binding 1 405 423 PF01473CW binding 1 525 538 APG07002 Seq ID no PFAM 158 domains APG07002 Seq IDSignal no PFAM modified 159 peptide domains (APG07002.1) removedAPG07020 Seq ID PF03945 Endotoxin N 84 248 160 APG07020 Seq ID Alternatestart PF03945 Endotoxin N 50 214 modified 161 (APG07020.1) APG07114 SeqID no PFAM 162 domains APG07114 Seq ID Signal no PFAM modified 163peptide domains (APG07114.1) removed APG07220 Seq ID no PFAM 164 domainsAPG07224 Seq ID PF03945 Endotoxin N 104 287 165 PF01473 CW binding 1 433446 PF01473 CW binding 1 517 530 APG07224 Seq ID 3′ Truncation PF03945Endotoxin N 104 288 modified 166 (APG07224.1) APG07444 Seq ID PF03318ETX MTX2 25 202 167 APG07445 Seq ID PF03945 Endotoxin N 75 251 168APG07445 Seq ID Alternate start PF03945 Endotoxin N 14 196 modified 169(APG07445.1) APG07470 Seq ID PF03945 Endotoxin N 1 162 170 APG07639 SeqID PF03318 ETX MTX2 23 247 171 APG07676 Seq ID no PFAM 172 domainsAPG07676 Seq ID Alternate start no PFAM modified 173 domains(APG07676.1) APG07682 Seq ID no PFAM 174 domains APG07682 Seq ID Signalno PFAM modified 175 peptide domains (APG07682.1) removed APG07738 SeqID PF03318 ETX MTX2 10 253 176 APG07738 Seq ID Alternate start PF03318ETX MTX2 13 251 modified 177 (APG07738.1) APG07780 Seq ID no PFAM 178domains APG07954 Seq ID PF03945 Endotoxin N 78 291 179 PF00555 EndotoxinM 299 523 PF03944 Endotoxin C 533 684 APG08029 Seq ID no PFAM 180domains APG08029 Seq ID Alternate start no PFAM modified 181 domains(APG08029.1) APG08138 Seq ID PF03318 ETX MTX2 39 280 182 APG08151 Seq IDPF06101 DUF946 19 271 183 APG08151 Seq ID Alternate start PF06101 DUF94611 261 modified 184 (APG08151.1) APG08509 Seq ID PF03318 ETX MTX2 82 289185 APG08509 Seq ID Alternate start PF03318 ETX MTX2 60 267 modified 186(APG08509.1) APG08607 Seq ID PF03945 Endotoxin N 181 378 187 PF01473 CWbinding 1 475 494 PF01473 CW binding 1 584 604 PF01473 CW binding 1 619635 APG08607 Seq ID Signal PF03945 Endotoxin N 80 277 modified 188peptide PF01473 CW binding 1 374 393 (APG08607.2) removed PF01473 CWbinding 1 483 503 PF01473 CW binding 1 518 534 Alternate start PF03945Endotoxin N 118 315 APG08607 Seq ID PF01473 CW binding 1 412 431modified 189 PF01473 CW binding 1 521 541 (APG08607.1) PF01473 CWbinding 1 556 572 APG08628 Seq ID no PFAM 190 domains APG08628 Seq IDAlternate start no PFAM modified 191 domains (APG08628.1) APG08780 SeqID PF01338 Bac thur toxin 2 193 192 APG08780 Seq ID Alternate startPF01338 Bac thur toxin 2 193 modified 193 (APG08780.1) APG08794 Seq IDPF03318 ETX MTX2 87 297 194 APG08794 Seq ID Signal PF03318 ETX MTX2 50266 modified 195 peptide (APG08794.1) removed APG09055 Seq ID PF03318ETX MTX2 78 305 196 APG09055 Seq ID Signal PF03318 ETX MTX2 46 272modified 197 peptide (APG09055.1) removed APG09096 Seq ID PF16403DUF5011 263 336 198 APG09096 Seq ID Signal PF16403 DUF5011 238 311modified 199 peptide (APG09096.1) removed APG09376 Seq ID PF03945Endotoxin N 75 288 200 PF00555 Endotoxin M 296 499 PF03944 Endotoxin C509 642 APG09376 Seq ID 3′ Truncation PF03945 Endotoxin N 75 288modified 201 PF00555 Endotoxin M 296 499 (APG09376.1) PF03944 EndotoxinC 509 641 APG09455 Seq ID PF03318 ETX MTX2 23 292 202 APG09455 Seq IDAlternate start PF03318 ETX MTX2 34 274 modified 203 (APG09455.1)APG09642 Seq ID PF03318 ETX MTX2 39 292 204 APG09642 Seq ID SignalPF03318 ETX MTX2 28 281 modified 205 peptide (APG09642.1) removedAPG09659 Seq ID no PFAM 206 domains APG09659 Seq ID Alternate start noPFAM modified 207 domains (APG09659.1) APG09717 Seq ID PF01338 Bac thurtoxin 1 199 208 APG09726 Seq ID no PFAM 209 domains APG09726 Seq IDSignal no PFAM modified 210 peptide domains (APG09726.1) removedAPG09735 Seq ID PF03318 ETX MTX2 107 356 211 APG09735 Seq ID SignalPF03318 ETX MTX2 80 330 modified 212 peptide (APG09735.1) removedAPG02067 Seq ID PF07029 CryBP1 74 224 CryBP1 213 (APG04834) APG00743 SeqID no PFAM 214 domains APG00743 Seq ID Signal no PFAM modified 215peptide domains (APG00743.1) removed APG01705 Seq ID no PFAM 216 domainsAPG01705 Seq ID Signal no PFAM modified 217 peptide domains (APG01705.1)removed APG01989 Seq ID PF01338 Bac thur toxin 2 199 218 APG02245 Seq IDno PFAM 219 domains APG02279 Seq ID PF03318 ETX MTX2 35 256 220 APG02279Seq ID Alternate start PF03318 ETX MTX2 28 249 modified 221 (APG02279.1)APG02429 Seq ID no PFAM 222 domains APG02674 Seq ID no PFAM 223 domainsAPG02674 Seq ID Signal no PFAM modified 224 peptide domains (APG02674.1)removed APG02768 Seq ID no PFAM 225 domains APG03040 Seq ID no PFAM 226domains APG03185 Seq ID no PFAM 227 domains APG03185 Seq ID Signal noPFAM modified 228 peptide domains (APG03185.1) removed APG03217 Seq IDno PFAM 229 domains APG03217 Seq ID Alternate start no PFAM modified 230domains (APG03217.1) APG03368 Seq ID no PFAM 231 domains APG03368 Seq IDSignal no PFAM modified 232 peptide domains (APG03368.1) removedAPG03662 Seq ID no PFAM 233 domains APG03662 Seq ID PF07029 CryBP1 37187 CryBP1 234 (APG07022) APG04224 Seq ID no PFAM 235 domains APG04224Seq ID Alternate start no PFAM modified 236 domains (APG04224.1)APG04226 Seq ID no PFAM 237 domains APG04226 Seq ID Signal no PFAMmodified 238 peptide domains (APG04226.1) removed APG04485 Seq ID noPFAM 239 domains APG04485 Seq ID Signal no PFAM modified 240 peptidedomains (APG04485.1) removed APG04643 Seq ID no PFAM 241 domainsAPG04643 Seq ID Signal no PFAM modified 242 peptide domains (APG04643.2)removed APG04643 Seq ID Alternate start no PFAM modified 243 domains(APG04643.1) APG05660 Seq ID PF03945 Endotoxin N 75 288 244 PF00555Endotoxin M 296 499 PF03944 Endotoxin C 509 642 APG05660 Seq ID 3′Truncation PF03945 Endotoxin N 75 288 modified 245 PF00555 Endotoxin M296 499 (APG05660.1) PF03944 Endotoxin C 509 641 APG05969 Seq ID no PFAM246 domains APG06528 Seq ID PF03945 Endotoxin N 79 291 247 PF00555Endotoxin M 299 502 PF03944 Endotoxin C 512 645 APG06528 Seq IDAlternate start PF03945 Endotoxin N 76 288 modified 248 PF00555Endotoxin M 296 499 (APG06528.1) PF03944 Endotoxin C 509 642 APG06528Seq ID no PFAM Split-Cry 249 domains C-term (APG09626) APG07049 Seq IDno PFAM 250 domains APG07049 Seq ID Signal no PFAM modified 251 peptidedomains (APG07049.1) removed APG07574 Seq ID no PFAM 252 domainsAPG07574 Seq ID Signal no PFAM modified 253 peptide domains (APG07574.1)removed APG08043 Seq ID PF03318 ETX MTX2 9 254 254 APG08043 Seq IDAlternate start PF03318 ETX MTX2 12 252 modified 255 (APG08043.1)APG08085 Seq ID no PFAM 256 domains APG08085 Seq ID Signal no PFAMmodified 257 peptide domains (APG08085.1) removed APG08225 Seq IDPF03318 ETX MTX2 119 352 258 APG08225 Seq ID Signal PF03318 ETX MTX2 90323 modified 259 peptide (APG08225.1) removed APG08241 Seq ID PF03318ETX MTX2 26 247 260 APG08411 Seq ID PF03318 ETX MTX2 60 322 261 APG08411Seq ID Signal PF03318 ETX MTX2 29 291 modified 262 peptide (APG08411.1)removed APG08718 Seq ID PF03318 ETX MTX2 21 247 263 APG08973 Seq IDPF03945 Endotoxin N 165 365 264 PF01473 CW binding 1 492 507 PF01473 CWbinding 1 524 543 APG08973 Seq ID Alternate start PF03945 Endotoxin N128 328 modified 265 PF01473 CW binding 1 455 470 (APG08973.1) PF01473CW binding 1 487 506 APG08990 Seq ID no PFAM 266 domains APG08990 Seq IDSignal no PFAM modified 267 peptide domains (APG08990.1) removedAPG09256 Seq ID no PFAM 268 domains APG09842 Seq ID no PFAM 269 domainsAPG02960 Seq ID no PFAM 270 domains APG02960 Seq ID Alternate start noPFAM modified 271 domains (APG02960.1) APG02960 Seq ID Signal no PFAMmodified 272 peptide domains (APG02960.2) removed APG01265.0 SEQ IDPF03318 Clostridium 46 280 273 epsilon toxin ETX/Bacillus mosquitocidaltoxin MTX2 APG03954.0 SEQ ID PF03318 Clostridium 155 298 274 epsilontoxin ETX/Bacillus mosquitocidal toxin MTX2 APG03954.1 SEQ ID SignalPF03318 Clostridium 126 269 275 peptide epsilon toxin removedETX/Bacillus mosquitocidal toxin MTX2 APG04375.0 SEQ ID No PFAM 276domains APG04375.1 SEQ ID Alternate start No PFAM 277 domains APG04900.0SEQ ID PF01473 Putative cell 363 375 278 wall binding repeat PF01473Putative cell 390 402 wall binding repeat PF01473 Putative cell 417 430wall binding repeat APG04900.1 279 Signal PF01473 Putative cell 325 337peptide wall binding removed repeat PF01473 Putative cell 352 364 wallbinding repeat PF01473 Putative cell 379 392 wall binding repeatAPG00770.0 280 PF01338 Bacillus 5 189 thuringiensis toxin APG01577.0 281no PFAM domains APG01797.0 282 no PFAM domains APG01987.0 283 PF01338Bacillus 59 192 thuringiensis toxin APG02248.0 284 no PFAM domainsAPG02443.0 285 no PFAM domains APG02756.0 286 no PFAM domains APG03017.0287 no PFAM domains APG03148.0 288 no PFAM domains APG03574.0 289PF06101 Plant protein 27 228 of unknown function (DUF946) PF05431Insecticidal 264 420 Crystal Toxin, P42 APG03574.1 290 Alternat startPF06101 Plant protein 17 218 of unknown function (DUF946) PF05431Insecticidal 254 410 Crystal Toxin, P42 APG05399.0 291 no PFAM domainsAPG05678.0 292 no PFAM domains APG05707.0 293 PF01338 Bacillus 63 192thuringiensis toxin APG06385.0 294 PF01338 Bacillus 23 229 thuringiensistoxin APG08372.0 295 no PFAM domains APG08372.1 296 Signal no PFAMpeptide domains removed APG09444.0 297 PF01338 Bacillus 6 192thuringiensis toxin APG09444.1 298 Alternate start PF01338 Bacillus 5192 thuringiensis toxin APG09446.0 299 no PFAM domains APG09857.0 300 noPFAM domains APG09892.0 301 no PFAM domains APG01172 302 PF03945Endotoxin N 85 287 PF03944 Endotoxin C 487 654 APG01172 303 Alternatestart PF03945 Endotoxin N 68 270 modified and 3′ PF03944 Endotoxin C 470636 (APG01172.2) Truncation APG01172 304 Alternate start PF03945Endotoxin N 68 270 modified PF03944 Endotoxin C 470 637 (APG01172.1)APG01172 305 no PFAM Split-Cry domains C-terminus (APG06502) APG01288306 PF03945 Endotoxin N 208 329 PF03944 Endotoxin C 572 710 APG05711 3073′ Truncation PF03945 Endotoxin N 82 301 PF03944 Endotoxin C 504 636APG05711 308 PF03945 Endotoxin N 82 301 modified PF03944 Endotoxin C 504635 (APG05711.1) APG01288 309 3′ Truncation PF03945 Endotoxin N 209 329modified PF03944 Endotoxin C 573 709 (APG01288.1)

Recombinant or synthetic nucleic acids encoding the pesticidalpolypeptides disclosed herein are also provided. Of particular interestare nucleic acid sequences that have been designed for expression in aplant of interest. That is, the nucleic acid sequence can be optimizedfor increased expression in a host plant. A pesticidal protein of theinvention can be back-translated to produce a nucleic acid comprisingcodons optimized for expression in a particular host, for example, acrop plant. In another embodiment, the polynucleotides encoding thepolypeptides provided herein may be optimized for increased expressionin the transformed plant. That is, the polynucleotides can besynthesized using plant-preferred codons for improved expression. See,for example, Campbell and Gowri (1990) Plant Physiol. 92:1-11 for adiscussion of host-preferred codon usage. Methods are available in theart for synthesizing plant-preferred genes. See, for example, U.S. Pat.Nos. 5,380,831, and 5,436,391, and Murray et al. (1989) Nucleic AcidsRes. 17:477-498, herein incorporated by reference. Expression of such acoding sequence by the transformed plant (e.g., dicot or monocot) willresult in the production of a pesticidal polypeptide and conferincreased resistance in the plant to a pest. Recombinant and syntheticnucleic acid molecules encoding the pesticidal proteins of the inventiondo not include the naturally occurring bacterial sequence encoding theprotein.

A “recombinant polynucleotide” or “recombinant nucleic acid” comprises acombination of two or more chemically linked nucleic acid segments whichare not found directly joined in nature. By “directly joined” isintended the two nucleic acid segments are immediately adjacent andjoined to one another by a chemical linkage. In specific embodiments,the recombinant polynucleotide comprises a polynucleotide of interest ora variant or fragment thereof such that an additional chemically linkednucleic acid segment is located either 5′, 3′ or internal to thepolynucleotide of interest. Alternatively, the chemically-linked nucleicacid segment of the recombinant polynucleotide can be formed by deletionof a sequence. The additional chemically linked nucleic acid segment orthe sequence deleted to join the linked nucleic acid segments can be ofany length, including for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20or greater nucleotides. Various methods for making such recombinantpolynucleotides include chemical synthesis or by the manipulation ofisolated segments of polynucleotides by genetic engineering techniques.In specific embodiments, the recombinant polynucleotide can comprise arecombinant DNA sequence or a recombinant RNA sequence. A “fragment of arecombinant polynucleotide or nucleic acid” comprises at least one of acombination of two or more chemically linked amino acid segments whichare not found directly joined in nature.

Fragments of a polynucleotide (RNA or DNA) may encode protein fragmentsthat retain activity. In specific embodiments, a fragment of arecombinant polynucleotide or a recombinant polynucleotide constructcomprises at least one junction of the two or more chemically linked oroperably linked nucleic acid segments which are not found directlyjoined in nature. A fragment of a polynucleotide that encodes abiologically active portion of a polypeptide that retains pesticidalactivity will encode at least 25, 30, 40, 50, 60, 70, 75, 80, 90, 100,110, 120, 125, 130, 140, 150, 160, 170, 175, 180, contiguous aminoacids, or up to the total number of amino acids present in a full-lengthpolypeptide as set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64,65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253,254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267,268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, and/or309. In specific embodiments, such polypeptide fragments are activefragment, and in still other embodiments, the polypeptide fragmentcomprises a recombinant polypeptide fragment. As used herein, a fragmentof a recombinant polypeptide comprises at least one of a combination oftwo or more chemically linked amino acid segments which are not founddirectly joined in nature.

By “variants” is intended to mean substantially similar sequences. Forpolynucleotides, a variant comprises a deletion and/or addition of oneor more nucleotides at one or more internal sites within the nativepolynucleotide and/or a substitution of one or more nucleotides at oneor more sites in the native polynucleotide. As used herein, a “native”polynucleotide or polypeptide comprises a naturally occurring nucleotidesequence or amino acid sequence, respectively.

Variants of a particular polynucleotide of the invention (i.e., thereference polynucleotide) can also be evaluated by comparison of thepercent sequence identity between the polypeptide encoded by a variantpolynucleotide and the polypeptide encoded by the referencepolynucleotide. Thus, for example, an isolated polynucleotide thatencodes a polypeptide with a given percent sequence identity to thepolypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50,51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103,104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117,118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131,132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145,146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159,160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173,174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187,188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201,202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215,216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228,229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242,243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256,257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270,271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284,285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298,299, 300, 301, 302, 303, 304, 305, 306, 307, 308, and/or 309 aredisclosed. Percent sequence identity between any two polypeptides can becalculated using sequence alignment programs and parameters describedelsewhere herein. Where any given pair of polynucleotides of theinvention is evaluated by comparison of the percent sequence identityshared by the two polypeptides they encode, the percent sequenceidentity between the two encoded polypeptides is at least about 40%,45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or moresequence identity to SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48,49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66,67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101,102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115,116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129,130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143,144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157,158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171,172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185,186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199,200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213,214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225, 226,227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254,255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268,269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282,283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, and/or 309.In other embodiments, the variant of the polynucleotide provided hereindiffers from the native sequence by at least 1, 2, 3, 4, 5, 6, 7, 8, 9,10 or more nucleotides.

Variant polynucleotide and proteins also encompass sequences andproteins derived from a mutagenic and recombinogenic procedure such asDNA shuffling. With such a procedure, one or more different pesticidalprotein disclosed herein (SEQ ID NO: 1-309) is manipulated to create anew pesticidal protein possessing the desired properties. In thismanner, libraries of recombinant polynucleotides are generated from apopulation of related sequence polynucleotides comprising sequenceregions that have substantial sequence identity and can be homologouslyrecombined in vitro or in vivo. For example, using this approach,sequence motifs encoding a domain of interest may be shuffled betweenthe pesticidial sequences provided herein and other known pesticidialgenes to obtain a new gene coding for a protein with an improvedproperty of interest, such as an increased Km in the case of an enzyme.Strategies for such DNA shuffling are known in the art. See, forexample, Stemmer (1994) Proc. Natl. Acad. Sci. USA 91:10747-10751;Stemmer (1994) Nature 370:389-391; Crameri et al. (1997) Nature Biotech.15:436-438; Moore et al. (1997) J. Mol. Biol. 272:336-347; Zhang et al.(1997) Proc. Natl. Acad. Sci. USA 94:4504-4509; Crameri et al. (1998)Nature 391:288-291; and U.S. Pat. Nos. 5,605,793 and 5,837,458. A“shuffled” nucleic acid is a nucleic acid produced by a shufflingprocedure such as any shuffling procedure set forth herein. Shufflednucleic acids are produced by recombining (physically or virtually) twoor more nucleic acids (or character strings), for example in anartificial, and optionally recursive, fashion. Generally, one or morescreening steps are used in shuffling processes to identify nucleicacids of interest; this screening step can be performed before or afterany recombination step. In some (but not all) shuffling embodiments, itis desirable to perform multiple rounds of recombination prior toselection to increase the diversity of the pool to be screened. Theoverall process of recombination and selection are optionally repeatedrecursively. Depending on context, shuffling can refer to an overallprocess of recombination and selection, or, alternately, can simplyrefer to the recombinational portions of the overall process.

In one embodiment, a method of obtaining a polynucleotide that encodesan improved polypeptide comprising pesticidal activity is provided,wherein the improved polypeptide has at least one improved property overany one of SEQ ID NOS: 1-309. Such methods can comprises (a) recombininga plurality of parental polynucleotides to produce a library ofrecombinant polynucleotides encoding recombinant pesticidalpolypeptides; (b) screening the library to identify a recombinantpolynucleotide that encodes an improved recombinant pesticidalpolypeptide that has an enhanced property improved over the parentalpolynucleotide; (c) recovering the recombinant polynucleotide thatencodes the improved recombinant pesticidal polypeptide identified in(b); and, (d) repeating steps (a), (b) and (c) using the recombinantpolynucleotide recovered in step (c) as one of the plurality of parentalpolynucleotides in repeated step (a).

iii. Sequence Comparisons

As used herein, the term “identity” or “percent identity” when used withrespect to a particular pair of aligned amino acid sequences, refers tothe percent amino acid sequence identity that is obtained by countingthe number of identical matches in the alignment and dividing suchnumber of identical matches by the length of the aligned sequences. Asused herein, the term “similarity” or “percent similarity” when usedwith respect to a particular pair of aligned amino acid sequences,refers to the sum of the scores that are obtained from a scoring matrixfor each amino acid pair in the alignment divided by the length of thealigned sequences.

Unless otherwise stated, identity and similarity will be calculated bythe Needleman-Wunsch global alignment and scoring algorithms (Needlemanand Wunsch (1970) J. Mol. Biol. 48(3):443-453) as implemented by the“needle” program, distributed as part of the EMBOSS software package(Rice, P. Longden, I. and Bleasby, A., EMBOSS: The European MolecularBiology Open Software Suite, 2000, Trends in Genetics 16, (6) pp276-277, versions 6.3.1 available from EMBnet atembnet.org/resource/emboss and emboss.sourceforge.net, among othersources) using default gap penalties and scoring matrices (EBLOSUM62 forprotein and EDNAFULL for DNA). Equivalent programs may also be used. By“equivalent program” is intended any sequence comparison program that,for any two sequences in question, generates an alignment havingidentical nucleotide residue matches and an identical percent sequenceidentity when compared to the corresponding alignment generated byneedle from EMBOSS version 6.3.1.

Additional mathematical algorithms are known in the art and can beutilized for the comparison of two sequences. See, for example, thealgorithm of Karlin and Altschul (1990) Proc. Natl. Acad. Sci. USA87:2264, modified as in Karlin and Altschul (1993) Proc. Natl. Acad.Sci. USA 90:5873-5877. Such an algorithm is incorporated into the BLASTprograms of Altschul et al. (1990) J. Mol. Biol. 215:403. BLASTnucleotide searches can be performed with the BLASTN program (nucleotidequery searched against nucleotide sequences) to obtain nucleotidesequences homologous to pesticidal-like nucleic acid molecules of theinvention, or with the BLASTX program (translated nucleotide querysearched against protein sequences) to obtain protein sequenceshomologous to pesticidal nucleic acid molecules of the invention. BLASTprotein searches can be performed with the BLASTP program (protein querysearched against protein sequences) to obtain amino acid sequenceshomologous to pesticidal protein molecules of the invention, or with theTBLASTN program (protein query searched against translated nucleotidesequences) to obtain nucleotide sequences homologous to pesticidalprotein molecules of the invention. To obtain gapped alignments forcomparison purposes, Gapped BLAST (in BLAST 2.0) can be utilized asdescribed in Altschul et al. (1997) Nucleic Acids Res. 25:3389.Alternatively, PSI-Blast can be used to perform an iterated search thatdetects distant relationships between molecules. See Altschul et al.(1997) supra. When utilizing BLAST, Gapped BLAST, and PSI-Blastprograms, the default parameters of the respective programs (e.g.,BLASTX and BLASTN) can be used. Alignment may also be performed manuallyby inspection.

Two sequences are “optimally aligned” when they are aligned forsimilarity scoring using a defined amino acid substitution matrix (e.g.,BLOSUM62), gap existence penalty and gap extension penalty so as toarrive at the highest score possible for that pair of sequences. Aminoacid substitution matrices and their use in quantifying the similaritybetween two sequences are well-known in the art and described, e.g., inDayhoff et al. (1978) “A model of evolutionary change in proteins.” In“Atlas of Protein Sequence and Structure,” Vol. 5, Suppl. 3 (ed. M. O.Dayhoff), pp. 345-352. Natl. Biomed. Res. Found., Washington, D.C. andHenikoff et al. (1992) Proc. Natl. Acad. Sci. USA 89:10915-10919. TheBLOSUM62 matrix is often used as a default scoring substitution matrixin sequence alignment protocols. The gap existence penalty is imposedfor the introduction of a single amino acid gap in one of the alignedsequences, and the gap extension penalty is imposed for each additionalempty amino acid position inserted into an already opened gap. Thealignment is defined by the amino acids positions of each sequence atwhich the alignment begins and ends, and optionally by the insertion ofa gap or multiple gaps in one or both sequences, so as to arrive at thehighest possible score. While optimal alignment and scoring can beaccomplished manually, the process is facilitated by the use of acomputer-implemented alignment algorithm, e.g., gapped BLAST 2.0,described in Altschul et al. (1997) Nucleic Acids Res. 25:3389-3402, andmade available to the public at the National Center for BiotechnologyInformation Website (www.ncbi.nlm.nih.gov). Optimal alignments,including multiple alignments, can be prepared using, e.g., PSI-BLAST,available through www.ncbi.nlm.nih.gov and described by Altschul et al.(1997) Nucleic Acids Res. 25:3389-3402.

With respect to an amino acid sequence that is optimally aligned with areference sequence, an amino acid residue “corresponds to” the positionin the reference sequence with which the residue is paired in thealignment. The “position” is denoted by a number that sequentiallyidentifies each amino acid in the reference sequence based on itsposition relative to the N-terminus. For example, in SEQ ID NO: 1position 1 is M, position 2 is A, position 3 is I, etc. When a testsequence is optimally aligned with SEQ ID NO: 1, a residue in the testsequence that aligns with the I at position 3 is said to “correspond toposition 3” of SEQ ID NO: 1. Owing to deletions, insertion, truncations,fusions, etc., that must be taken into account when determining anoptimal alignment, in general the amino acid residue number in a testsequence as determined by simply counting from the N-terminal will notnecessarily be the same as the number of its corresponding position inthe reference sequence. For example, in a case where there is a deletionin an aligned test sequence, there will be no amino acid thatcorresponds to a position in the reference sequence at the site ofdeletion. Where there is an insertion in an aligned reference sequence,that insertion will not correspond to any amino acid position in thereference sequence. In the case of truncations or fusions there can bestretches of amino acids in either the reference or aligned sequencethat do not correspond to any amino acid in the corresponding sequence.

iv. Antibodies

Antibodies to the polypeptides of the present invention, or to variantsor fragments thereof, are also encompassed. Methods for producingantibodies are well known in the art (see, for example, Harlow and Lane(1988) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory,Cold Spring Harbor, N.Y.; and U.S. Pat. No. 4,196,265). These antibodiescan be used in kits for the detection and isolation of toxinpolypeptides. Thus, this disclosure provides kits comprising antibodiesthat specifically bind to the polypeptides described herein, including,for example, polypeptides having the sequence of SEQ ID NOs: 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110,111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124,125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138,139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152,153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166,167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180,181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194,195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208,209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221,222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235,236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249,250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263,264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291,292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305,306, 307, 308, and/or 309.

II. Pests

The compositions and methods provided herein are useful against avariety of pests. “Pests” includes but is not limited to, insects,fungi, bacteria, nematodes, acarids, protozoan pathogens,animal-parasitic liver flukes, and the like. Pests of particularinterest are insect pests, particularly insect pests that causesignificant damage to agricultural plants. Insect pests include insectsselected from the orders Coleoptera, Diptera, Hymenoptera, Lepidoptera,Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera,Isoptera, Anoplura, Siphonaptera, Trichoptera, or nematodes. Innon-limiting embodiments, the insect pest comprises Western cornrootworm, Diabrotica virgifera virgifera; Fall armyworm, Spodopterafrugiperda; Colorado potato beetle, Leptinotarsa decemlineata; Cornearworm, Helicoverpa zea (in North America same species attacks cottonand called cotton bollworm); European corn borer, Ostrinia nubilalis;Black cutworm, Agrotis ipsilon; Diamondback moth, Plutella xylostella;Velvetbean caterpillar, Anticarsia gemmatalis; Southwestern corn borer,Diatraea grandiosella; Cotton bollworm, Helicoverpa armigera (foundother than USA in rest of the world); Southern green stinkbug, Nezaraviridula; Green stinkbug, Chinavia halaris; Brown marmorated stinkbug,Halyomorpha halys; and Brown stinbug, Euschistus servus, Euschistusheros (Neotropical brown stink bug OR soy stink bug); Piezodorusguildinii (red-banded stink bug); Dichelops melacanthus (no common name)and/or Dichelops furcatus (no common name); an aphid, such as a soybeanaphid. In other embodiments, the pest comprises a nematode including,but not limited to, Meloidogyne hapla (Northern root-knot nematode);Meloidogyne enterolobii, Meloidogyne arenaria (peanut root-knotnematode); and Meloidogyne javanica.

The term “insect pests” as used herein refers to insects and othersimilar pests such as, for example, those of the order Acari including,but not limited to, mites and ticks. Insect pests of the presentinvention include, but are not limited to, insects of the orderLepidoptera, e.g. Achoroia grisella, Acleris gloverana, Acleris variana,Adoxophyes orana, Agrotis ipsilon, Alabama argillacea, Alsophilapometaria, Amyelois transitella, Anagasta kuehniella, Anarsialineatella, Anisota senatoria, Antheraea pernyi, Anticarsia gemmatalis,Archips sp., Argyrotaenia sp., Athetis mindara, Bombyx mori, Bucculatrixthurberiella, Cadra cautella, Choristoneura sp., Cochylls hospes, Coliaseurytheme, Corcyra cephalonica, Cydia latiferreanus, Cydia pomonella,Datana integerrima, Dendrolimus sibericus, Desmiafeneralis, Diaphaniahyalinata, Diaphania nitidalis, Diatraea grandiosella, Diatraeasaccharalis, Ennomos subsignaria, Eoreuma loftini, Esphestia elutella,Erannis tilaria, Estigmene acrea, Eulia salubricola, Eupocoelliaambiguella, Eupoecilia ambiguella, Euproctis chrysorrhoea, Euxoamessoria, Galleria mellonella, Grapholita molesta, Harrisina americana,Helicoverpa subflexa, Helicoverpa zea, Heliothis virescens, Hemileucaoliviae, Homoeosoma electellum, Hyphantia cunea, Keiferialycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellarialugubrosa, Leucoma salicis, Lobesia botrana, Loxostege sticticalis,Lymantria dispar, Macalla thyrisalis, Malacosoma sp., Mamestrabrassicae, Mamestra configurata, Manduca quinquemaculata, Manduca sexta,Maruca testulalis, Melanchra picta, Operophtera brumata, Orgyia sp.,Ostrinia nubilalis, Paleacrita vernata, Papilio cresphontes,Pectinophora gossypiella, Phryganidia californica, Phyllonorycterblancardella, Pieris napi, Pieris rapae, Plathypena scabra, Platynotaflouendana, Platynota stultana, Platyptilia carduidactyla, Plodiainterpunctella, Plutella xylostella, Pontia protodice, Pseudaletiaunipuncta, Pseudoplasia includens, Sabulodes aegrotata, Schizuraconcinna, Sitotroga cerealella, Spilonta ocellana, Spodoptera sp.,Thaurnstopoea pityocampa, Tinsola bisselliella, Trichoplusia hi, Udearubigalis, Xylomyges curtails, and Yponomeuta padella.

Insect pests also include insects selected from the orders Diptera,Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera,Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera,Coleoptera.

Insect pests of the invention for the major crops include, but are notlimited to: Maize: Ostrinia nubilalis, European corn borer; Agrotisipsilon, black cutworm; Helicoverpa zeae, corn earworm; Spodopterafrugiperda, fall armyworm; Diatraea grandiosella, southwestern cornborer; Elasmopalpus lignosellus, lesser cornstalk borer; Diatraeasaccharalis, surgarcane borer; western corn rootworm, e.g., Diabroticavirgifera virgifera; northern corn rootworm, e.g., Diabroticalongicornis barberi; southern corn rootworm, e.g., Diabroticaundecimpunctata howardi; Melanotus spp., wireworms; Cyclocephalaborealis, northern masked chafer (white grub); Cyclocephala immaculata,southern masked chafer (white grub); Popillia japonica, Japanese beetle;Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maizebillbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis,corn root aphid; Euschistus heros (Neotropical brown stink bug OR soystink bug); Piezodorus guildinii (red-banded stink bug); Dichelopsmelacanthus (no common name); Dichelops furcatus (no common name);Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum,redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper;Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blotchleafminer; Anaphothrips obscrurus, grass thrips; Solenopsis milesta,thief ant; Tetranychus urticae, two spotted spider mite; Sorghum: Chilopartellus, sorghum borer; Spodoptera frugiperda, fall armyworm;Helicoverpa zea, corn earworm; Elasmopalpus lignosellus, leser cornstalkborer; Feltia subterranea, granulate cutworm; Phyllophaga crinita, whitegrub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus,cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle;Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leafaphid; Sipha flava, yellow sugarcane aphid; chinch bug, e.g., Blissusleucopterus leucopterus; Contarinia sorghicola, sorghum midge;Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae,two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm;Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lessercornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpuslignosellus, lesser cornstalk borer; Oulema melanopus, cereal leafbeetle; Hypera punctata, clover leaf weevil; southern corn rootworm,e.g., Diabrotica undecimpunctata howardi; Russian wheat aphid;Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid;Melanoplus femurrubrum, redlegged grasshopper; Melanoplusdifferentialis, differential grasshopper; Melanoplus sanguinipes,migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosismosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemyacoarctata, wheat bulb fly; Frankliniella fusca, tobacco thrips; Cephuscinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower:Cylindrocupturus adspersus, sunflower stem weevil; Smicronyx fulus, redsunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil;Suleima helianthana, sunflower bud moth; Homoeosoma electellum,sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrusgibbosus, carrot beetle; Neolasioptera murtfeldtiana, sunflower seedmidge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea,cotton bollworm; Spodoptera exigua, beet armyworm; Pectinophoragossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphisgossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper;Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris,tarnished plant bug; Melanoplus femurrubrum, redlegged grasshopper;Melanoplus differentialis, differential grasshopper; Thrips tabaci,onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychuscinnabarinus, carmine spider mite; Tetranychus urticae, two-spottedspider mite; Rice: Diatraea saccharalis, sugarcane borer; Spodopterafrugiperda, fall armyworm; Helicoverpa zea, corn earworm; Colaspisbrunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil;Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhoper;chinch bug, e.g., Blissus leucopterus leucopterus; Acrosternum hilare,green stink bug; Soybean: Pseudoplusia includens, soybean looper;Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, greencloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon,black cutworm; Spodoptera exigua, beet armyworm; Heliothis virescens,tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis,Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae,potato leafhopper; Acrosternum hilare, green stink bug; Melanoplusfemurrubrum, redlegged grasshopper; Melanoplus differentialis,differential grasshopper; Hylemya platura, seedcorn maggot; Sericothripsvariabilis, soybean thrips; Thrips tabaci, onion thrips; Tetranychusturkestani, strawberry spider mite; Tetranychus urticae, two-spottedspider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotisipsilon, black cutworm; Schizaphis graminum, greenbug; chinch bug, e.g.,Blissus leucopterus leucopterus; Acrosternum hilare, green stink bug;Euschistus servus, brown stink bug; Jylemya platura, seedcorn maggot;Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite;Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotretacruciferae, crucifer flea beetle; Phyllotreta striolata, striped fleabeetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethesaeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus,Meligethes nigrescens, Meligethes canadianus, and Meligethesviridescens; Potato: Leptinotarsa decemlineata, Colorado potato beetle.

The methods and compositions provided herein may be effective againstHemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis,Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthopscompestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltisnotatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis,Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus,Poecilocapsus lineatus, Blissus leucopterus, Nysius ericae, Nysiusraphanus, Euschistus servus, Nezara viridula, Eurygaster, Coreidae,Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, and Cimicidae. Pestsof interest also include Araecerus fasciculatus, coffee bean weevil;Acanthoscelides obtectus, bean weevil; Bruchus rufmanus, broadbeanweevil; Bruchus pisorum, pea weevil; Zabrotes subfasciatus, Mexican beanweevil; Diabrotica balteata, banded cucumber beetle; Cerotomatrifurcata, bean leaf beetle; Diabrotica virgifera, Mexican cornrootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis,sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomusquadrigibbus, apple curculio; Sternechus paludatus, bean stalk weevil;Hypera brunnipennis, Egyptian alfalfa weevil; Sitophilus granaries,granary weevil; Craponius inaequalis, grape curculio; Sitophiluszeamais, maize weevil; Conotrachelus nenuphar, plum curculio; Euscepespostfaciatus, West Indian sweet potato weevil; Maladera castanea,Asiatic garden beetle; Rhizotrogus majalis, European chafer;Macrodactylus subspinosus, rose chafer; Tribolium confusum, confusedflour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, redflour beetle; Tenebrio molitor, yellow mealworm.

Nematodes include parasitic nematodes such as root-knot, cyst, andlesion nematodes, including Heterodera spp., Meloidogyne spp., andGlobodera spp.; particularly members of the cyst nematodes, including,but not limited to, Heterodera glycines (soybean cyst nematode);Heterodera schachtii (beet cyst nematode); Heterodera avenae (cerealcyst nematode); and Globodera rostochiensis and Globodera pailida(potato cyst nematodes). Lesion nematodes include Pratylenchus spp.

Insect pests may be tested for pesticidal activity of compositions ofthe invention in early developmental stages, e.g., as larvae or otherimmature forms. The insects may be reared in total darkness at fromabout 20° C. to about 30° C. and from about 30% to about 70% relativehumidity. Bioassays may be performed as described in Czapla and Lang(1990) J. Econ. Entomol. 83 (6): 2480-2485. See, also the experimentalsection herein.

III. Expression Cassettes

Polynucleotides encoding the pesticidal proteins provided herein can beprovided in expression cassettes for expression in an organism ofinterest. The cassette will include 5′ and 3′ regulatory sequencesoperably linked to a polynucleotide encoding a pesticidal polypeptideprovided herein that allows for expression of the polynucleotide. Thecassette may additionally contain at least one additional gene orgenetic element to be cotransformed into the organism. Where additionalgenes or elements are included, the components are operably linked.Alternatively, the additional gene(s) or element(s) can be provided onmultiple expression cassettes. Such an expression cassette is providedwith a plurality of restriction sites and/or recombination sites forinsertion of the polynucleotides to be under the transcriptionalregulation of the regulatory regions. The expression cassette mayadditionally contain a selectable marker gene.

The expression cassette will include in the 5′-3′ direction oftranscription, a transcriptional and translational initiation region(i.e., a promoter), a pesticidal polynucleotide of the invention, and atranscriptional and translational termination region (i.e., terminationregion) functional in the organism of interest, i.e., a plant orbacteria. The promoters of the invention are capable of directing ordriving expression of a coding sequence in a host cell. The regulatoryregions (i.e., promoters, transcriptional regulatory regions, andtranslational termination regions) may be endogenous or heterologous tothe host cell or to each other. As used herein, “heterologous” inreference to a sequence is a sequence that originates from a foreignspecies, or, if from the same species, is substantially modified fromits native form in composition and/or genomic locus by deliberate humanintervention. As used herein, a chimeric gene comprises a codingsequence operably linked to a transcription initiation region that isheterologous to the coding sequence.

Convenient termination regions are available from the Ti-plasmid of A.tumefaciens, such as the octopine synthase and nopaline synthasetermination regions. See also Guerineau et al. (1991) Mol. Gen. Genet.262:141-144; Proudfoot (1991) Cell 64:671-674; Sanfacon et al. (1991)Genes Dev. 5:141-149; Mogen et al. (1990) Plant Cell 2:1261-1272; Munroeet al. (1990) Gene 91:151-158; Ballas et al. (1989) Nucleic Acids Res.17:7891-7903; and Joshi et al. (1987) Nucleic Acids Res. 15:9627-9639.

Additional regulatory signals include, but are not limited to,transcriptional initiation start sites, operators, activators,enhancers, other regulatory elements, ribosomal binding sites, aninitiation codon, termination signals, and the like. See, for example,U.S. Pat. Nos. 5,039,523 and 4,853,331; EPO 0480762A2; Sambrook et al.(1992) Molecular Cloning: A Laboratory Manual, ed. Maniatis et al. (ColdSpring Harbor Laboratory Press, Cold Spring Harbor, N.Y.), hereinafter“Sambrook 11”; Davis et al., eds. (1980) Advanced Bacterial Genetics(Cold Spring Harbor Laboratory Press), Cold Spring Harbor, N.Y., and thereferences cited therein.

In preparing the expression cassette, the various DNA fragments may bemanipulated, so as to provide for the DNA sequences in the properorientation and, as appropriate, in the proper reading frame. Towardthis end, adapters or linkers may be employed to join the DNA fragmentsor other manipulations may be involved to provide for convenientrestriction sites, removal of superfluous DNA, removal of restrictionsites, or the like. For this purpose, in vitro mutagenesis, primerrepair, restriction, annealing, resubstitutions, e.g., transitions andtransversions, may be involved.

A number of promoters can be used in the practice of the invention. Thepromoters can be selected based on the desired outcome. The nucleicacids can be combined with constitutive, inducible, tissue-preferred, orother promoters for expression in the organism of interest. See, forexample, promoters set forth in WO 99/43838 and in U.S. Pat. Nos.8,575,425; 7,790,846; 8,147,856; 8,586832; 7,772,369; 7,534,939;6,072,050; 5,659,026; 5,608,149; 5,608,144; 5,604,121; 5,569,597;5,466,785; 5,399,680; 5,268,463; 5,608,142; and 6,177,611; hereinincorporated by reference.

For expression in plants, constitutive promoters also include CaMV 35Spromoter (Odell et al. (1985) Nature 313:810-812); rice actin (McElroyet al. (1990) Plant Cell 2:163-171); ubiquitin (Christensen et al.(1989) Plant Mol. Biol. 12:619-632 and Christensen et al. (1992) PlantMol. Biol. 18:675-689); pEMU (Last et al. (1991) Theor. Appl. Genet.81:581-588); MAS (Velten et al. (1984) EMBO J. 3:2723-2730). Induciblepromoters include those that drive expression of pathogenesis-relatedproteins (PR proteins), which are induced following infection by apathogen. See, for example, Redolfi et al. (1983) Neth. J. Plant Pathol.89:245-254; Uknes et al. (1992) Plant Cell 4:645-656; and Van Loon(1985) Plant Mol. Virol. 4:111-116; and WO 99/43819, herein incorporatedby reference. Promoters that are expressed locally at or near the siteof pathogen infection may also be used (Marineau et al. (1987) PlantMol. Biol. 9:335-342; Matton et al. (1989) Molecular Plant-MicrobeInteractions 2:325-331; Somsisch et al. (1986) Proc. Natl. Acad. Sci.USA 83:2427-2430; Somsisch et al. (1988) Mol. Gen. Genet. 2:93-98; andYang (1996) Proc. Natl. Acad. Sci. USA 93:14972-14977; Chen et al.(1996) Plant J. 10:955-966; Zhang et al. (1994) Proc. Natl. Acad. Sci.USA 91:2507-2511; Warner et al. (1993) Plant J. 3:191-201; Siebertz etal. (1989) Plant Cell 1:961-968; Cordero et al. (1992) Physiol. Mol.Plant Path. 41:189-200; U.S. Pat. No. 5,750,386 (nematode-inducible);and the references cited therein).

Wound-inducible promoters may be used in the constructions of theinvention. Such wound-inducible promoters include pin II promoter (Ryan(1990) Ann. Rev. Phytopath. 28:425-449; Duan et al. (1996) NatureBiotechnology 14:494-498); wun1 and wun2 (U.S. Pat. No. 5,428,148); win1and win2 (Stanford et al. (1989) Mol. Gen. Genet. 215:200-208); systemin(McGurl et al. (1992) Science 225:1570-1573); WIP1 (Rohmeier et al.(1993) Plant Mol. Biol. 22:783-792; Eckelkamp et al. (1993) FEBS Letters323:73-76); MPI gene (Corderok et al. (1994) Plant J. 6(2):141-150); andthe like, herein incorporated by reference.

Tissue-preferred promoters for use in the invention include those setforth in Yamamoto et al. (1997) Plant J. 12(2):255-265; Kawamata et al.(1997) Plant Cell Physiol. 38(7):792-803; Hansen et al. (1997) Mol. GenGenet. 254(3):337-343; Russell et al. (1997) Transgenic Res.6(2):157-168; Rinehart et al. (1996) Plant Physiol. 112(3):1331-1341;Van Camp et al. (1996) Plant Physiol. 112(2):525-535; Canevascini et al.(1996) Plant Physiol. 112(2):513-524; Yamamoto et al. (1994) Plant CellPhysiol. 35(5):773-778; Lam (1994) Results Probl. Cell Differ.20:181-196; Orozco et al. (1993) Plant Mol Biol. 23(6):1129-1138;Matsuoka et al. (1993) Proc Natl. Acad. Sci. USA 90(20):9586-9590; andGuevara-Garcia et al. (1993) Plant J. 4(3):495-505.

Leaf-preferred promoters include those set forth in Yamamoto et al.(1997) Plant J. 12(2):255-265; Kwon et al. (1994) Plant Physiol.105:357-67; Yamamoto et al. (1994) Plant Cell Physiol. 35(5):773-778;Gotor et al. (1993) Plant J. 3:509-18; Orozco et al. (1993) Plant Mol.Biol. 23(6):1129-1138; and Matsuoka et al. (1993) Proc. Natl. Acad. Sci.USA 90(20):9586-9590.

Root-preferred promoters are known and include those in Hire et al.(1992) Plant Mol. Biol. 20(2):207-218 (soybean root-specific glutaminesynthetase gene); Keller and Baumgartner (1991) Plant Cell3(10):1051-1061 (root-specific control element); Sanger et al. (1990)Plant Mol. Biol. 14(3):433-443 (mannopine synthase (MAS) gene ofAgrobacterium tumefaciens); and Miao et al. (1991) Plant Cell 3(1):11-22(cytosolic glutamine synthetase (GS)); Bogusz et al. (1990) Plant Cell2(7):633-641; Leach and Aoyagi (1991) Plant Science (Limerick)79(1):69-76 (rolC and rolD); Teeri et al. (1989) EMBO J. 8(2):343-350;Kuster et al. (1995) Plant Mol. Biol. 29(4):759-772 (the VfENOD-GRP3gene promoter); and, Capana et al. (1994) Plant Mol. Biol. 25(4):681-691(rolB promoter). See also U.S. Pat. Nos. 5,837,876; 5,750,386;5,633,363; 5,459,252; 5,401,836; 5,110,732; and 5,023,179.

“Seed-preferred” promoters include both “seed-specific” promoters (thosepromoters active during seed development such as promoters of seedstorage proteins) as well as “seed-germinating” promoters (thosepromoters active during seed germination). See Thompson et al. (1989)BioEssays 10:108. Seed-preferred promoters include, but are not limitedto, Cim1 (cytokinin-induced message); cZ19B1 (maize 19 kDa zein); mi1ps(myo-inositol-1-phosphate synthase) (see WO 00/11177 and U.S. Pat. No.6,225,529). Gamma-zein is an endosperm-specific promoter. Globulin 1(Glb-1) is a representative embryo-specific promoter. For dicots,seed-specific promoters include, but are not limited to, beanβ-phaseolin, napin, β-conglycinin, soybean lectin, cruciferin, and thelike. For monocots, seed-specific promoters include, but are not limitedto, maize 15 kDa zein, 22 kDa zein, 27 kDa zein, gamma-zein, waxy,shrunken 1, shrunken 2, Globulin 1, etc. See also WO 00/12733, whereseed-preferred promoters from end1 and end2 genes are disclosed.

For expression in a bacterial host, promoters that function in bacteriaare well-known in the art. Such promoters include any of the knowncrystal protein gene promoters, including the promoters of any of thepesticidal proteins of the invention, and promoters specific for B.thuringiensis sigma factors. Alternatively, mutagenized or recombinantcrystal protein-encoding gene promoters may be recombinantly engineeredand used to promote expression of the novel gene segments disclosedherein.

The expression cassette can also comprise a selectable marker gene forthe selection of transformed cells. Selectable marker genes are utilizedfor the selection of transformed cells or tissues. Marker genes includegenes encoding antibiotic resistance, such as those encoding neomycinphosphotransferase II (NEO) and hygromycin phosphotransferase (HPT), aswell as genes conferring resistance to herbicidal compounds, such asglufosinate ammonium, bromoxynil, imidazolinones, and2,4-dichlorophenoxyacetate (2,4-D). Additional selectable markers areknown and any can be used. See, for example, U.S. Provisionalapplication 62/094,697, filed on Dec. 19, 2014, and U.S. ProvisionalApplication 62/189,505, filed Jul. 7, 2015, both of which are hereinincorporated by reference in their entirety, which discloses glufosinateresistance sequences that can be employed as selectable markers. See,for example, PCT/US2015/066648, filed on Dec. 18, 2015, hereinincorporated by reference in its entirety, which discloses glufosinateresistance sequences that can be employed as selectable markers.

IV. Methods, Host Cells and Plant Cells

As indicated, DNA constructs comprising nucleotide sequences encodingthe pesticidal proteins or active variants or fragment thereof can beused to transform plants of interest or other organisms of interest.Methods for transformation involve introducing a nucleotide constructinto a plant. By “introducing” is intended to introduce the nucleotideconstruct to the plant or other host cell in such a manner that theconstruct gains access to the interior of a cell of the plant or hostcell. The methods of the invention do not require a particular methodfor introducing a nucleotide construct to a plant or host cell, onlythat the nucleotide construct gains access to the interior of at leastone cell of the plant or the host organism. Methods for introducingnucleotide constructs into plants and other host cells are known in theart including, but not limited to, stable transformation methods,transient transformation methods, and virus-mediated methods.

The methods result in a transformed organisms, such as a plant,including whole plants, as well as plant organs (e.g., leaves, stems,roots, etc.), seeds, plant cells, propagules, embryos and progeny of thesame. Plant cells can be differentiated or undifferentiated (e.g.callus, suspension culture cells, protoplasts, leaf cells, root cells,phloem cells, pollen).

“Transgenic plants” or “transformed plants” or “stably transformed”plants or cells or tissues refers to plants that have incorporated orintegrated a polynucleotide encoding at least one pesticidal polypeptideof the invention. It is recognized that other exogenous or endogenousnucleic acid sequences or DNA fragments may also be incorporated intothe plant cell. Agrobacterium- and biolistic-mediated transformationremain the two predominantly employed approaches. However,transformation may be performed by infection, transfection,microinjection, electroporation, microprojection, biolistics or particlebombardment, electroporation, silica/carbon fibers, ultrasound mediated,PEG mediated, calcium phosphate co-precipitation, polycation DMSOtechnique, DEAE dextran procedure, Agro and viral mediated(Caulimoriviruses, Geminiviruses, RNA plant viruses), liposome mediatedand the like.

Transformation protocols as well as protocols for introducingpolypeptides or polynucleotide sequences into plants may vary dependingon the type of plant or plant cell, i.e., monocot or dicot, targeted fortransformation. Methods for transformation are known in the art andinclude those set forth in U.S. Pat. Nos. 8,575,425; 7,692,068;8,802,934; 7,541,517; each of which is herein incorporated by reference.See, also, Rakoczy-Trojanowska, M. (2002) Cell Mol Biol Lett. 7:849-858;Jones et al. (2005) Plant Methods 1:5; Rivera et al. (2012) Physics ofLife Reviews 9:308-345; Bartlett et al. (2008) Plant Methods 4:1-12;Bates, G. W. (1999) Methods in Molecular Biology 111:359-366; Binns andThomashow (1988) Annual Reviews in Microbiology 42:575-606; Christou, P.(1992) The Plant Journal 2:275-281; Christou, P. (1995) Euphytica85:13-27; Tzfira et al. (2004) TRENDS in Genetics 20:375-383; Yao et al.(2006) Journal of Experimental Botany 57:3737-3746; Zupan and Zambryski(1995) Plant Physiology 107:1041-1047; Jones et al. (2005) Plant Methods1:5.

Transformation may result in stable or transient incorporation of thenucleic acid into the cell. “Stable transformation” is intended to meanthat the nucleotide construct introduced into a host cell integratesinto the genome of the host cell and is capable of being inherited bythe progeny thereof “Transient transformation” is intended to mean thata polynucleotide is introduced into the host cell and does not integrateinto the genome of the host cell.

Methods for transformation of chloroplasts are known in the art. See,for example, Svab et al. (1990) Proc. Nail. Acad. Sci. USA 87:8526-8530;Svab and Maliga (1993) Proc. Natl. Acad. Sci. USA 90:913-917; Svab andMaliga (1993) EMBO J. 12:601-606. The method relies on particle gundelivery of DNA containing a selectable marker and targeting of the DNAto the plastid genome through homologous recombination. Additionally,plastid transformation can be accomplished by transactivation of asilent plastid-borne transgene by tissue-preferred expression of anuclear-encoded and plastid-directed RNA polymerase. Such a system hasbeen reported in McBride et al. (1994) Proc. Natl. Acad. Sci. USA91:7301-7305.

The cells that have been transformed may be grown into plants inaccordance with conventional ways. See, for example, McCormick et al.(1986) Plant Cell Reports 5:81-84. These plants may then be grown, andeither pollinated with the same transformed strain or different strains,and the resulting hybrid having constitutive expression of the desiredphenotypic characteristic identified. Two or more generations may begrown to ensure that expression of the desired phenotypic characteristicis stably maintained and inherited and then seeds harvested to ensureexpression of the desired phenotypic characteristic has been achieved.In this manner, the present invention provides transformed seed (alsoreferred to as “transgenic seed”) having a nucleotide construct of theinvention, for example, an expression cassette of the invention, stablyincorporated into their genome.

In specific embodiments, the sequences provide herein can be targeted tospecific cite within the genome of the host cell or plant cell. Suchmethods include, but are not limited to, meganucleases designed againstthe plant genomic sequence of interest (D'Halluin et al. 2013 PlantBiotechnol J); CRISPR-Cas9, TALENs, and other technologies for preciseediting of genomes (Feng, et al. Cell Research 23:1229-1232, 2013,Podevin, et al. Trends Biotechnology, online publication, 2013, Wei etal., J Gen Genomics, 2013, Zhang et al (2013) WO 2013/026740); Cre-loxsite-specific recombination (Dale et al. (1995) Plant J 7:649-659;Lyznik, et al. (2007) Transgenic Plant J 1:1-9; FLP-FRT recombination(Li et al. (2009) Plant Physiol 151:1087-1095); Bxb1-mediatedintegration (Yau et al. Plant J (2011) 701:147-166); zinc-fingermediated integration (Wright et al. (2005) Plant J 44:693-705); Cai etal. (2009) Plant Mol Biol 69:699-709); and homologous recombination(Lieberman-Lazarovich and Levy (2011) Methods Mol Biol 701: 51-65);Puchta (2002) Plant Mol Biol 48:173-182).

The sequence provided herein may be used for transformation of any plantspecies, including, but not limited to, monocots and dicots. Examples ofplants of interest include, but are not limited to, corn (maize),sorghum, wheat, sunflower, tomato, crucifers, peppers, potato, cotton,rice, soybean, sugarbeet, sugarcane, tobacco, barley, and oilseed rape,Brassica sp., alfalfa, rye, millet, safflower, peanuts, sweet potato,cassaya, coffee, coconut, pineapple, citrus trees, cocoa, tea, banana,avocado, fig, guava, mango, olive, papaya, cashew, macadamia, almond,oats, vegetables, ornamentals, and conifers.

Vegetables include, but are not limited to, tomatoes, lettuce, greenbeans, lima beans, peas, and members of the genus Curcumis such ascucumber, cantaloupe, and musk melon. Ornamentals include, but are notlimited to, azalea, hydrangea, hibiscus, roses, tulips, daffodils,petunias, carnation, poinsettia, and chrysanthemum. Preferably, plantsof the present invention are crop plants (for example, maize, sorghum,wheat, sunflower, tomato, crucifers, peppers, potato, cotton, rice,soybean, sugarbeet, sugarcane, tobacco, barley, oilseed rape, etc.).

As used herein, the term plant includes plant cells, plant protoplasts,plant cell tissue cultures from which plants can be regenerated, plantcalli, plant clumps, and plant cells that are intact in plants or partsof plants such as embryos, pollen, ovules, seeds, leaves, flowers,branches, fruit, kernels, ears, cobs, husks, stalks, roots, root tips,anthers, and the like. Grain is intended to mean the mature seedproduced by commercial growers for purposes other than growing orreproducing the species. Progeny, variants, and mutants of theregenerated plants are also included within the scope of the invention,provided that these parts comprise the introduced polynucleotides.Further provided is a processed plant product or byproduct that retainsthe sequences disclosed herein, including for example, soymeal.

In another embodiment, the genes encoding the pesticidal proteins can beused to transform insect pathogenic organisms. Such organisms includebaculoviruses, fungi, protozoa, bacteria, and nematodes. Microorganismhosts that are known to occupy the “phytosphere” (phylloplane,phyllosphere, rhizosphere, and/or rhizoplana) of one or more crops ofinterest may be selected. These microorganisms are selected so as to becapable of successfully competing in the particular environment with thewild-type microorganisms, provide for stable maintenance and expressionof the gene expressing the pesticidal protein, and desirably, providefor improved protection of the pesticide from environmental degradationand inactivation.

Such microorganisms include archaea, bacteria, algae, and fungi. Ofparticular interest are microorganisms such as bacteria, e.g., Bacillus,Pseudomonas, Erwinia, Serratia, Klebsiella, Xanthomonas, Streptomyces,Rhizobium, Rhodopseudomonas, Methylius, Agrobacterium, Acetobacter,Lactobacillus, Arthrobacter, Azotobacter, Leuconostoc, and Alcaligenes.Fungi include yeast, e.g., Saccharomyces, Cryptococcus, Kluyveromyces,Sporobolomyces, Rhodotorula, and Aureobasidium. Of particular interestare such phytosphere bacterial species as Pseudomonas syringae,Pseudomonas aeruginosa, Pseudomonas fluorescens, Serratia marcescens,Acetobacter xylinum, Agrobacteria, Rhodopseudomonas spheroides,Xanthomonas campestris, Rhizobium melioti, Alcaligenes entrophus,Clavibacter xyli and Azotobacter vinlandir and phytosphere yeast speciessuch as Rhodotorula rubra, R. glutinis, R. marina, R. aurantiaca,Cryptococcus albidus, C. diffluens, C. laurentii, Saccharomyces rosei,S. pretoriensis, S. cerevisiae, Sporobolomyces rosues, S. odorus,Kluyveromyces veronae, Aureobasidium pollulans, Bacillus thuringiensis,Escherichia coli, Bacillus subtilis, and the like.

Illustrative prokaryotes, both Gram-negative and gram-positive, includeEnterobacteriaceae, such as Escherichia, Erwinia, Shigella, Salmonella,and Proteus; Bacillaceae; Rhizobiceae, such as Rhizobium; Spirillaceae,such as photobacterium, Zymomonas, Serratia, Aeromonas, Vibrio,Desulfovibrio, Spirillum; Lactobacillaceae; Pseudomonadaceae, such asPseudomonas and Acetobacter; Azotobacteraceae and Nitrobacteraceae.Fungi include Phycomycetes and Ascomycetes, e.g., yeast, such asSaccharomyces and Schizosaccharomyces; and Basidiomycetes yeast, such asRhodotorula, Aureobasidium, Sporobolomyces, and the like.

Genes encoding pesticidal proteins can be introduced by means ofelectrotransformation, PEG induced transformation, heat shock,transduction, conjugation, and the like. Specifically, genes encodingthe pesticidal proteins can be cloned into a shuttle vector, forexample, pHT3101 (Lerecius et al. (1989) FEMS Microbiol. Letts. 60:211-218. The shuttle vector pHT3101 containing the coding sequence forthe particular pesticidal protein gene can, for example, be transformedinto the root-colonizing Bacillus by means of electroporation (Lereciuset al. (1989) FEMS Microbiol. Letts. 60: 211-218).

Expression systems can be designed so that pesticidal proteins aresecreted outside the cytoplasm of gram-negative bacteria by fusing anappropriate signal peptide to the amino-terminal end of the pesticidalprotein. Signal peptides recognized by E. coli include the OmpA protein(Ghrayeb et al. (1984) EMBO J, 3: 2437-2442).

Pesticidal proteins and active variants thereof can be fermented in abacterial host and the resulting bacteria processed and used as amicrobial spray in the same manner that Bacillus thuringiensis strainshave been used as insecticidal sprays. In the case of a pesticidalprotein(s) that is secreted from Bacillus, the secretion signal isremoved or mutated using procedures known in the art. Such mutationsand/or deletions prevent secretion of the pesticidal protein(s) into thegrowth medium during the fermentation process. The pesticidal proteinsare retained within the cell, and the cells are then processed to yieldthe encapsulated pesticidal proteins.

Alternatively, the pesticidal proteins are produced by introducingheterologous genes into a cellular host. Expression of the heterologousgene results, directly or indirectly, in the intracellular productionand maintenance of the pesticide. These cells are then treated underconditions that prolong the activity of the toxin produced in the cellwhen the cell is applied to the environment of target pest(s). Theresulting product retains the toxicity of the toxin. These naturallyencapsulated pesticidal proteins may then be formulated in accordancewith conventional techniques for application to the environment hostinga target pest, e.g., soil, water, and foliage of plants. See, forexample U.S. Pat. No. 6,468,523 and U.S. Publication No. 20050138685,and the references cited therein. In the present invention, atransformed microorganism (which includes whole organisms, cells,spore(s), pesticidal protein(s), pesticidal component(s), pest-impactingcomponent(s), mutant(s), living or dead cells and cell components,including mixtures of living and dead cells and cell components, andincluding broken cells and cell components) or an isolated pesticidalprotein can be formulated with an acceptable carrier into a pesticidalor agricultural composition(s) that is, for example, a suspension, asolution, an emulsion, a dusting powder, a dispersible granule, awettable powder, and an emulsifiable concentrate, an aerosol, animpregnated granule, an adjuvant, a coatable paste, and alsoencapsulations in, for example, polymer substances.

Agricultural compositions may comprise a polypeptide, a recombinogenicpolypeptide or a variant or fragment thereof, as disclosed herein. Theagricultural composition disclosed herein may be applied to theenvironment of a plant or an area of cultivation, or applied to theplant, plant part, plant cell, or seed.

Such compositions disclosed above may be obtained by the addition of asurface-active agent, an inert carrier, a preservative, a humectant, afeeding stimulant, an attractant, an encapsulating agent, a binder, anemulsifier, a dye, a UV protectant, a buffer, a flow agent orfertilizers, micronutrient donors, or other preparations that influenceplant growth. One or more agrochemicals including, but not limited to,herbicides, insecticides, fungicides, bactericides, nematicides,molluscicides, acaracides, plant growth regulators, harvest aids, andfertilizers, can be combined with carriers, surfactants or adjuvantscustomarily employed in the art of formulation or other components tofacilitate product handling and application for particular target pests.Suitable carriers and adjuvants can be solid or liquid and correspond tothe substances ordinarily employed in formulation technology, e.g.,natural or regenerated mineral substances, solvents, dispersants,wetting agents, tackifiers, binders, or fertilizers. The activeingredients of the present invention are normally applied in the form ofcompositions and can be applied to the crop area, plant, or seed to betreated. For example, the compositions of the present invention may beapplied to grain in preparation for or during storage in a grain bin orsilo, etc. The compositions of the present invention may be appliedsimultaneously or in succession with other compounds. Methods ofapplying an active ingredient of the present invention or anagrochemical composition of the present invention that contains at leastone of the pesticidal proteins produced by the bacterial strains of thepresent invention include, but are not limited to, foliar application,seed coating, and soil application. The number of applications and therate of application depend on the intensity of infestation by thecorresponding pest.

Suitable surface-active agents include, but are not limited to, anioniccompounds such as a carboxylate of, for example, a metal; a carboxylateof a long chain fatty acid; an N-acylsarcosinate; mono or di-esters ofphosphoric acid with fatty alcohol ethoxylates or salts of such esters;fatty alcohol sulfates such as sodium dodecyl sulfate, sodium octadecylsulfate or sodium cetyl sulfate; ethoxylated fatty alcohol sulfates;ethoxylated alkylphenol sulfates; lignin sulfonates; petroleumsulfonates; alkyl aryl sulfonates such as alkyl-benzene sulfonates orlower alkylnaphtalene sulfonates, e.g., butyl-naphthalene sulfonate;salts of sulfonated naphthalene-formaldehyde condensates; salts ofsulfonated phenol-formaldehyde condensates; more complex sulfonates suchas the amide sulfonates, e.g., the sulfonated condensation product ofoleic acid and N-methyl taurine; or the dialkyl sulfosuccinates, e.g.,the sodium sulfonate of dioctyl succinate. Non-ionic agents includecondensation products of fatty acid esters, fatty alcohols, fatty acidamides or fatty-alkyl- or alkenyl-substituted phenols with ethyleneoxide, fatty esters of polyhydric alcohol ethers, e.g., sorbitan fattyacid esters, condensation products of such esters with ethylene oxide,e.g., polyoxyethylene sorbitar fatty acid esters, block copolymers ofethylene oxide and propylene oxide, acetylenic glycols such as2,4,7,9-tetraethyl-5-decyn-4,7-diol, or ethoxylated acetylenic glycols.Examples of a cationic surface-active agent include, for instance, analiphatic mono-, di-, or polyamine such as an acetate, naphthenate oroleate; or oxygen-containing amine such as an amine oxide ofpolyoxyethylene alkylamine; an amide-linked amine prepared by thecondensation of a carboxylic acid with a di- or polyamine; or aquaternary ammonium salt.

Examples of inert materials include but are not limited to inorganicminerals such as kaolin, phyllosilicates, carbonates, sulfates,phosphates, or botanical materials such as cork, powdered corncobs,peanut hulls, rice hulls, and walnut shells.

The compositions of the present invention can be in a suitable form fordirect application or as a concentrate of primary composition thatrequires dilution with a suitable quantity of water or other diluantbefore application. The pesticidal concentration will vary dependingupon the nature of the particular formulation, specifically, whether itis a concentrate or to be used directly. The composition contains 1 to98% of a solid or liquid inert carrier, and 0 to 50% or 0.1 to 50% of asurfactant. These compositions will be administered at the labeled ratefor the commercial product, for example, about 0.01 lb-5.0 lb. per acrewhen in dry form and at about 0.01 pts.-10 pts. per acre when in liquidform.

In a further embodiment, the compositions, as well as the transformedmicroorganisms and pesticidal proteins, provided herein can be treatedprior to formulation to prolong the pesticidal activity when applied tothe environment of a target pest as long as the pretreatment is notdeleterious to the pesticidal activity. Such treatment can be bychemical and/or physical means as long as the treatment does notdeleteriously affect the properties of the composition(s). Examples ofchemical reagents include but are not limited to halogenating agents;aldehydes such as formaldehyde and glutaraldehyde; anti-infectives, suchas zephiran chloride; alcohols, such as isopropanol and ethanol; andhistological fixatives, such as Bouin's fixative and Helly's fixative(see, for example, Humason (1967) Animal Tissue Techniques (W.H. Freemanand Co.).

In one aspect, pests may be killed or reduced in numbers in a given areaby application of the pesticidal proteins provided herein to the area.Alternatively, the pesticidal proteins may be prophylactically appliedto an environmental area to prevent infestation by a susceptible pest.Preferably the pest ingests, or is contacted with, apesticidally-effective amount of the polypeptide. By“pesticidally-effective amount” is intended an amount of the pesticidethat is able to bring about death to at least one pest, or to noticeablyreduce pest growth, feeding, or normal physiological development. Thisamount will vary depending on such factors as, for example, the specifictarget pests to be controlled, the specific environment, location,plant, crop, or agricultural site to be treated, the environmentalconditions, and the method, rate, concentration, stability, and quantityof application of the pesticidally-effective polypeptide composition.The formulations or compositions may also vary with respect to climaticconditions, environmental considerations, and/or frequency ofapplication and/or severity of pest infestation.

The active ingredients are normally applied in the form of compositionsand can be applied to the crop area, plant, or seed to be treated.Methods are therefore provided for providing to a plant, plant cell,seed, plant part or an area of cultivation, an effective amount of theagricultural composition comprising the polypeptide, recombinogenicpolypeptide or an active variant or fragment thereof. By “effectiveamount” is intended an amount of a protein or composition has pesticidalactivity that is sufficient to kill or control the pest or result in anoticeable reduction in pest growth, feeding, or normal physiologicaldevelopment. Such decreases in numbers, pest growth, feeding or normaldevelopment can comprise any statistically significant decrease,including, for example a decrease of about 5%, 10%, 15%, 20%, 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 85%, 90%, 95% or greater.For example, the compositions may be applied to grain in preparation foror during storage in a grain bin or silo, etc. The compositions may beapplied simultaneously or in succession with other compounds. Methods ofapplying an active ingredient or an agrochemical composition comprisingat least one of the polypeptides, recombinogenic polypeptides orvariants or fragments thereof as disclosed herein, include but are notlimited to, foliar application, seed coating, and soil application.

Methods for increasing plant yield are provided. The methods compriseproviding a plant or plant cell expressing a polynucleotide encoding thepesticidal polypeptide sequence disclosed herein and growing the plantor a seed thereof in a field infested with (or susceptible toinfestation by) a pest against which said polypeptide has pesticidalactivity. In some embodiments, the polypeptide has pesticidal activityagainst a lepidopteran, coleopteran, dipteran, hemipteran, or nematodepest, and said field is infested with a lepidopteran, hemipteran,coleopteran, dipteran, or nematode pest. As defined herein, the “yield”of the plant refers to the quality and/or quantity of biomass producedby the plant. By “biomass” is intended any measured plant product. Anincrease in biomass production is any improvement in the yield of themeasured plant product. Increasing plant yield has several commercialapplications. For example, increasing plant leaf biomass may increasethe yield of leafy vegetables for human or animal consumption.Additionally, increasing leaf biomass can be used to increase productionof plant-derived pharmaceutical or industrial products. An increase inyield can comprise any statistically significant increase including, butnot limited to, at least a 1% increase, at least a 3% increase, at leasta 5% increase, at least a 10% increase, at least a 20% increase, atleast a 30%, at least a 50%, at least a 70%, at least a 100% or agreater increase in yield compared to a plant not expressing thepesticidal sequence. In specific methods, plant yield is increased as aresult of improved pest resistance of a plant expressing a pesticidalprotein disclosed herein. Expression of the pesticidal protein resultsin a reduced ability of a pest to infest or feed.

The plants can also be treated with one or more chemical compositions,including one or more herbicide, insecticides, or fungicides.

Non-limiting embodiments include:

1. An isolated polypeptide having insecticidal activity, comprising

(a) a polypeptide comprising an amino acid sequence selected from thegroup consisting of sequences set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61,62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223,224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279,280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293,294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307,308, and/or 309; or

(b) a polypeptide comprising an amino acid sequence having at least thepercent sequence identity set forth in Table 1 to an amino acid sequenceselected from the group consisting of sequences set forth in SEQ ID NOs:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38,39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56,57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108,109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122,123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136,137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150,151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164,165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178,179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192,193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206,207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 218, 219,220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233,234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247,248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261,262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275,276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289,290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303,304, 305, 306, 307, 308, and/or 309.

2. The polypeptide of embodiment 1, wherein said polypeptide comprisesthe amino acid sequence set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98,99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223,224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279,280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293,294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307,308, and/or 309.

3. A composition comprising the polypeptide of embodiments 1 or 2.

4. The polypeptide of embodiment 2, further comprising heterologousamino acid sequences.

5. A recombinant nucleic acid molecule that encodes the polypeptide ofembodiment 1, wherein said recombinant nucleic acid molecule is not thenaturally occurring sequence encoding said polypeptide.

6. The recombinant nucleic acid of embodiment 5, wherein said nucleicacid molecule is a synthetic sequence that has been designed forexpression in a plant.

7. The recombinant nucleic acid molecule of embodiment 6, wherein saidnucleic acid molecule is operably linked to a promoter capable ofdirecting expression in a plant cell.

8. The recombinant nucleic acid molecule of embodiment 5, wherein saidnucleic acid molecule is operably linked to a promoter capable ofdirecting expression in a bacteria.

9. A host cell that contains the recombinant nucleic acid molecule ofembodiment 8.

10. The host cell of embodiment 9, wherein said host cell is a bacterialhost cell.

11. A DNA construct comprising a promoter that drives expression in aplant cell operably linked to a recombinant nucleic acid moleculecomprising

(a) a nucleotide sequence that encodes a polypeptide comprising theamino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211,212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,and/or 309; or,

(b) a nucleotide sequence that encodes a polypeptide comprising an aminoacid sequence having at least the percent sequence identity set forth inTable 1 to an amino acid sequence selected from the group consisting ofsequences set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253,254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267,268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, and/or309.

12. The DNA construct of embodiment 11, wherein said nucleotide sequenceis a synthetic DNA sequence that has been designed for expression in aplant.

13. A vector comprising the DNA construct of embodiment 11.

14. A host cell that contains the DNA construct of embodiment 11 or 12or the vector of embodiment 13.

15. The host cell of embodiment 14, wherein the host cell is a plantcell.

16. A transgenic plant comprising the host cell of embodiment 15.

17. A composition comprising the host cell of embodiment 10.

18. The composition of embodiment 17, wherein said composition isselected from the group consisting of a powder, dust, pellet, granule,spray, emulsion, colloid, and solution.

19. The composition of embodiment 17, wherein said composition comprisesfrom about 1% to about 99% by weight of said polypeptide.

20. A method for controlling a pest population comprising contactingsaid population with a pesticidal-effective amount of the composition ofembodiment 17.

21. A method for killing a pest population comprising contacting saidpopulation with a pesticidal-effective amount of the composition ofembodiment 17.

22. A method for producing a polypeptide with pesticidal activity,comprising culturing the host cell of embodiment 9 under conditions inwhich the nucleic acid molecule encoding the polypeptide is expressed.

23. A plant having stably incorporated into its genome a DNA constructcomprising a nucleotide sequence that encodes a protein havingpesticidal activity, wherein said nucleotide sequence comprise

(a) a nucleotide sequence that encodes a polypeptide comprising theamino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211,212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,and/or 309; or,

(b) a nucleotide sequence that encodes a polypeptide comprising an aminoacid sequence having at least the percent sequence identity set forth inTable 1 to an amino acid sequence selected from the group consisting ofsequences set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253,254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267,268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, and/or309.

24. A transgenic seed of the plant of embodiment 23.

25. A method for protecting a plant from an insect pest, comprisingexpressing in a plant or cell thereof a nucleotide sequence that encodesa pesticidal polypeptide, wherein said nucleotide sequence comprising

(a) a nucleotide sequence that encodes a polypeptide comprising theamino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211,212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,and/or 309; or,

(b) a nucleotide sequence that encodes a polypeptide comprising an aminoacid sequence having at least the percent sequence identity set forth inTable 1 to an amino acid sequence selected from the group consisting ofsequences set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253,254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267,268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, and/or309.

26. The method of embodiment 25, wherein said plant produces apesticidal polypeptide having pesticidal against a lepidopteran orcoleopteran pest.

27. A method for increasing yield in a plant comprising growing in afield a plant or seed thereof having stably incorporated into its genomea DNA construct comprising a promoter that drives expression in a plantoperably linked to a nucleotide sequence that encodes a pesticidalpolypeptide, wherein said nucleotide sequence comprises

(a) a nucleotide sequence that encodes a polypeptide comprising theamino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211,212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,and/or 309; or,

(b) a nucleotide sequence that encodes a polypeptide comprising an aminoacid sequence having at least the percent sequence identity set forth inTable 1 to an amino acid sequence selected from the group consisting ofsequences set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253,254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267,268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, and/or309.

28. A method of obtaining a polynucleotide that encodes an improvedpolypeptide comprising pesticidal activity is provided, wherein theimproved polypeptide has at least one improved property over any one ofSEQ ID NOS: 1-279 comprising:

(a) recombining a plurality of parental polynucleotides comprising SEQID NO: 1-279 or an active variant or fragment thereof to produce alibrary of recombinant polynucleotides encoding recombinant pesticidalpolypeptides;

(b) screening the library to identify a recombinant polynucleotide thatencodes an improved recombinant pesticidal polypeptide that has anenhanced property improved over the parental polynucleotide;

(c) recovering the recombinant polynucleotide that encodes the improvedrecombinant pesticidal polypeptide identified in (b); and

(d) repeating steps (a), (b) and (c) using the recombinantpolynucleotide recovered in step (c) as one of the plurality of parentalpolynucleotides in repeated step (a).

EXAMPLES

The following examples are offered by way of illustration and not by wayof limitation.

Example 1: Discovery of Novel Genes by Sequencing and DNA Analysis

Microbial cultures were grown in liquid culture in standard laboratorymedia. Cultures were grown to saturation (16 to 24 hours) before DNApreparation. DNA was extracted from bacterial cells by detergent lysis,followed by binding to a silica matrix and washing with an ethanolbuffer. Purified DNA was eluted from the silica matrix with a mildlyalkaline aqueous buffer.

DNA for sequencing was tested for purity and concentration byspectrophotometry. Sequencing libraries were prepared using the NexteraXT library preparation kit according to the manufacturer's protocol.Sequence data was generated on a HiSeq 2000 according to the IlluminaHiSeq 2000 System User Guide protocol.

Sequencing reads were assembled into draft genomes using the CLC BioAssembly Cell software package. Following assembly, gene calls were madeby several methods and resulting gene sequences were interrogated toidentify novel homologs of pesticidal genes. Novel genes were identifiedby BLAST, by domain composition, and by pairwise alignment versus atarget set of pesticidal genes. A summary of such sequences is set forthin Table 1.

Genes identified in the homology search were amplified from bacterialDNA by PCR and cloned into bacterial expression vectors containing fusedin-frame purification tags. Cloned genes were expressed in E. coli andpurified by column chromatography. Purified proteins were assessed ininsect diet bioassay studies to identify active proteins.

Example 2. Heterologous Expression in E. coli

Each open reading frame set forth in SEQ ID NOS: 1-309 (or an activevariant or fragment thereof) is cloned into an E. coli expression vectorcontaining a maltose binding protein (pMBP). The expression vector istransformed into BL21*RIPL. An LB culture supplemented withcarbenicillin is inoculated with a single colony and grown overnight at37° C. using 0.5% of the overnight culture, a fresh culture isinoculated and grown to logarithmic phase at 37° C. The culture isinduced using 250 mM IPTG for 18 hours at 16° C. The cells are pelletedand resuspended in 10 mM Tris pH 7.4 and 150 mM NaCl supplemented withprotease inhibitors. The protein expression is evaluated by SDS-PAGE.

Example 3. Pesticidal Activity Against Coleopteran and Lepidoptera

Protein Expression: Each sequence set forth in SEQ ID NOS: 1-309 (or anactive variant or fragment thereof) is expressed in E. coli as describedin Example 2. 400 mL of LB is inoculated and grown to an OD600 of 0.6.The culture is induced with 0.25 mM IPTG overnight at 16° C. The cellsare spun down and the cell pellet is resuspend in 5 mL of buffer. Theresuspension is sonicated for 2 min on ice.

Bioassay: Fall army worm (FAW), corn ear worm (CEW), European corn borer(ECB) southwestern corn borer (SWCB) and diamond backed moth (DBM) eggsare purchased from a commercial insectary (Benzon Research Inc.,Carlisle, Pa.). The FAW, CEW, ECB and BCW eggs are incubated to thepoint that eclosion would occur within 12 hrs of the assay setup. SWCBand DBM are introduced to the assay as neonate larvae. Assays arecarried out in 24-well trays containing multispecies lepidopteran diet(SOUTHLAND PRODUCTS INCORPORATED, Lake Village, Ark.). Samples of thesonicated lysate are applied to the surface of the diet (diet overlay)and allowed to evaporate and soak into the diet. For CEW, FAW, BCW, ECBand SWCB, a 125 μl of sonicated lysate is added to the diet surface anddried. For DBM, 50 μl of a 1:2 dilution of sonicated lysate was added tothe diet surface. The bioassay plates are sealed with a plate sealingfilm vented with pin holes. The plates are incubated at 26° C. at 65% RHon a 16:8 day:night cycle in a Percival for 5 days. The assays areassessed for level of mortality, growth inhibition and feedinginhibition.

For the western corn rootworm bioassay, the protein construct/lysate isevaluated in an insect bioassay by dispensing 60 μl volume on the topsurface of diet in well/s of 24-well plate (Cellstar, 24-well, GreinerBio One) and allowed to dry. Each well contains 500 μl diet (Marrone etal., 1985). Fifteen to twenty neonate larvae are introduced in each wellusing a fine tip paint brush and the plate is covered with membrane(Viewseal, Greiner Bio One). The bioassay is stored at ambienttemperature and scored for mortality, and/or growth/feeding inhibitionat day 4.

For Colorado Potato Beetle (CPB) a cork bore size No. 8 leaf disk isexcised from potato leaf and is dipped in the protein construct/lysateuntil thoroughly wet and placed on top of filter disk (Millipore, glassfiber filter, 13 mm). Sixty μl dH2O is added to each filter disk andplaced in each well of 24-well plate (Cellstar, 24-well, Greiner BioOne). The leaf disk is allowed to dry and five to seven first instarlarvae are introduced in each well using a fine tip paint brush. Theplate is covered with membrane (Viewseal, Greiner Bio One) and smallhole is punctured in each well of the membrane. The construct isevaluated with four replicates, and scored for mortality and leaf damageon day 3.

Example 4. Pesticidal Activity Against Hemipteran

Protein Expression: Each of the sequences set forth in SEQ ID NOS: 1-309(or an active variant or fragment thereof) is expressed in E. coli asdescribed in Example 2. 400 mL of LB is inoculated and grown to an OD600of 0.6. The culture is induced with 0.25 mM IPTG overnight at 16° C. Thecells are spun down and the cell pellet is re-suspend in 5 mL of buffer.The resuspension is sonicated for 2 min on ice.

Second instar SGSB are obtained from a commercial insectary (BenzonResearch Inc., Carlisle, Pa.). A 50% v/v ratio of sonicated lysatesample to 20% sucrose is employed in the bioassay. Stretched parafilm isused as a feeding membrane to expose the SGSB to the diet/samplemixture. The plates are incubated at 25° C.:21° C., 16:8 day:night cycleat 65% RH for 5 days. Mortality is scored for each sample.

Example 5. Transformation of Soybean

DNA constructs comprising each of SEQ ID NOS: 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211,212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224,225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266,267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280,281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,and/or 309, or active variants or fragments of any thereof, operablylinked to a promoter active in a plant are cloned into transformationvectors and introduced into Agrobacterium as described in U.S.Provisional Application No. 62/094,782, filed Dec. 19, 2015, hereinincorporated by reference in its entirety.

Four days prior to inoculation, several loops of Agrobacterium arestreaked to a fresh plate of YEP* medium supplemented with theappropriate antibiotics** (spectinomycin, chloramphenicol andkanamycin). Bacteria are grown for two days in the dark at 28 C. Aftertwo days, several loops of bacteria are transferred to 3 ml of YEPliquid medium with antibiotics in a 125 ml Erlenmeyer flask. Flasks areplaced on a rotary shaker at 250 RPM at 28° C. overnight. One day beforeinoculation, 2-3 ml of the overnight culture were transferred to 125 mlof YEP with antibiotics in a 500 ml Erlenmeyer flask. Flasks are placedon a rotary shaker at 250 RPM at 28° C. overnight.

Prior to inoculation, the OD of the bacterial culture is checked at OD620. An OD of 0.8-1.0 indicates that the culture is in log phase. Theculture is centrifuged at 4000 RPM for 10 minutes in Oakridge tubes. Thesupernatant is discarded and the pellet is re-suspended in a volume ofSoybean Infection Medium (SI) to achieve the desired OD. The culturesare held with periodic mixing until needed for inoculation.

Two or three days prior to inoculation, soybean seeds are surfacesterilized using chlorine gas. In a fume hood, a petri dish with seedsis place in a bell jar with the lid off 1.75 ml of 12 N HCl is slowlyadded to 100 ml of bleach in a 250 ml Erlenmeyer flask inside the belljar. The lid is immediately placed on top of the bell jar. Seeds areallowed to sterilize for 14-16 hours (overnight). The top is removedfrom the bell jar and the lid of the petri dish is replaced. The petridish with the surface sterilized is then opened in a laminar flow foraround 30 minutes to disperse any remaining chlorine gas.

Seeds are imbibed with either sterile DI water or soybean infectionmedium (SI) for 1-2 days. Twenty to 30 seeds are covered with liquid ina 100×25 mm petri dish and incubated in the dark at 24 C. Afterimbibition, non-germinating seeds are discarded.

Cotyledonary explants is processed on a sterile paper plate with sterilefilter paper dampened using SI medium employing the methods of U.S. Pat.No. 7,473,822, herein incorporated by reference.

Typically, 16-20 cotyledons are inoculated per treatment. The SI mediumused for holding the explants is discarded and replaced with 25 ml ofAgrobacterium culture (OD 620=0.8-20). After all explants are submerged,the inoculation is carried out for 30 minutes with periodic swirling ofthe dish. After 30 minutes, the Agrobacterium culture is removed.

Co-cultivation plates is prepared by overlaying one piece of sterilepaper onto Soybean Co-cultivation Medium (SCC). Without blotting, theinoculated cotyledons is cultured adaxial side down on the filter paper.Around 20 explants can be cultured on each plate. The plates are sealedwith Parafilm and cultured at 24 C and around 120 umoles m-2s-1 (in aPercival incubator) for 4-5 days.

After co-cultivation, the cotyledons are washed 3 times in 25 ml ofSoybean Wash Medium with 200 mg/l of cefotaxime and timentin. Thecotyledons are blotted on sterile filter paper and then transferred toSoybean Shoot Induction Medium (SSI). The nodal end of the explant isdepressed slightly into the medium with distal end kept above thesurface at about 45 degrees. No more than 10 explants are cultured oneach plate. The plates are wrapped with Micropore tape and cultured inthe Percival at 24° C. and around 120 moles m-2s-1.

The explants are transferred to fresh SSI medium after 14 days. Emergingshoots from the shoot apex and cotyledonary node are discarded. Shootinduction is continued for another 14 days under the same conditions.

After 4 weeks of shoot induction, the cotyledon is separated from thenodal end and a parallel cut is made underneath the area of shootinduction (shoot pad). The area of the parallel cut is placed on SoybeanShoot Elongation Medium (SSE) and the explants cultured in the Percivalat 24° C. and around 120 moles m-2s-1. This step is repeated every twoweeks for up to 8 weeks as long as shoots continue to elongate.

When shoots reach a length of 2-3 cm, they are transferred to SoybeanRooting Medium (SR) in a Plantcon vessel and incubated under the sameconditions for 2 weeks or until roots reach a length of around 3-4 cm.After this, plants are transferred to soil.

Note, all media mentioned for soybean transformation are found in Paz etal. (2010) Agrobacterium-mediated transformation of soybean and recoveryof transgenic soybean plants; Plant Transformation Facility of IowaState University, which is herein incorporated by reference in itsentirety. (See, agron-www.agron.iastate.edu/ptf/protocol/Soybean.pdf.)

Example 6. Transformation of Maize

Maize ears are best collected 8-12 days after pollination. Embryos areisolated from the ears, and those embryos 0.8-1.5 mm in size arepreferred for use in transformation. Embryos are plated scutellumside-up on a suitable incubation media, such as DN62A5S media (3.98 g/LN6 Salts; 1 mL/L (of 1000.times. Stock) N6 Vitamins; 800 mg/LL-Asparagine; 100 mg/L Myo-inositol; 1.4 g/L L-Proline; 100 mg/LCasamino acids; 50 g/L sucrose; 1 mL/L (of 1 mg/mL Stock) 2,4-D).However, media and salts other than DN62A5S are suitable and are knownin the art. Embryos are incubated overnight at 25° C. in the dark.However, it is not necessary per se to incubate the embryos overnight.

The resulting explants are transferred to mesh squares (30-40 perplate), transferred onto osmotic media for about 30-45 minutes, thentransferred to a beaming plate (see, for example, PCT Publication No.WO/01/38514 and U.S. Pat. No. 5,240,842). DNA constructs designed toexpress the any one of SEQ ID NO: 1-309 or an active variant or fragmentthereof in plant cells are accelerated into plant tissue using anaerosol beam accelerator, using conditions essentially as described inPCT Publication No. WO/01/38514. After beaming, embryos are incubatedfor about 30 min on osmotic media, and placed onto incubation mediaovernight at 25° C. in the dark. To avoid unduly damaging beamedexplants, they are incubated for at least 24 hours prior to transfer torecovery media. Embryos are then spread onto recovery period media, forabout 5 days, 25° C. in the dark, then transferred to a selection media.Explants are incubated in selection media for up to eight weeks,depending on the nature and characteristics of the particular selectionutilized. After the selection period, the resulting callus istransferred to embryo maturation media, until the formation of maturesomatic embryos is observed. The resulting mature somatic embryos arethen placed under low light, and the process of regeneration isinitiated by methods known in the art. The resulting shoots are allowedto root on rooting media, and the resulting plants are transferred tonursery pots and propagated as transgenic plants.

Example 7. Pesticidal Activity Against Nematodes

A. Heterodera Glycine's (Soybean Cyst Nematode) In Vitro Assay

Soybean Cyst Nematodes are dispensed into a 96 well assay plate with atotal volume of 100 uls and 100 J2 per well. The protein of interest asset forth in any one of SEQ ID NOS: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239,240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253,254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267,268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281,282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295,296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, and/or309, or an active variant or fragment of any thereof, is dispensed intothe wells and held at room temperature for assessment. Finally, the 96well plate containing the SCN J2 is analyzed for motility. Data isreported as % inhibition as compared to the controls. Hits are definedas greater or equal to 70% inhibition.

B. Heterodera Glycine's (Soybean Cyst Nematode) On-Plant Assay

Soybean plants expressing one or more of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61,62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223,224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279,280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293,294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307,308, and/or 309, or an active variant or fragment of any thereof, aregenerated as described elsewhere herein. A 3-week-old soybean cutting isinoculated with 5000 SCN eggs per plant. This infection is held for 70days and then harvested for counting of SCN cyst that has developed onthe plant. Data is reported as % inhibition as compared to the controls.Hits are defined as greater or equal to 90% inhibition.

C. Meloidogyne incognita (Root-Knot Nematode) In Vitro Assay

Root-Knot Nematodes are dispensed into a 96 well assay plate with atotal volume of 100 uls and 100 J2 per well. The protein of interestcomprising any one of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48,49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66,67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101,102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115,116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129,130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143,144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157,158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171,172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185,186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199,200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213,214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225, 226,227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254,255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268,269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282,283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, and/or 309,or an active variant or fragment of any thereof, is dispensed into thewells and held at room temperature for assessment. Finally, the 96 wellplate containing the RKN J2 is analyzed for motility. Data is reportedas % inhibition as compared to the controls. Hits are defined as greateror equal to 70% inhibition.

D. Meloidogyne incognita (Root-Knot Nematode) On-Plant Assay

Soybean plants expressing one or more of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61,62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223,224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279,280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293,294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307,308, and/or 309, or an active variant or fragment of any thereof, aregenerated as described elsewhere herein. A 3-week-old soybean isinoculated with 5000 RKN eggs per plant. This infection is held for 70days and then harvested for counting of RKN eggs that have developed inthe plant. Data is reported as % inhibition as compared to the controls.Hits are defined as greater or equal to 90% inhibition.

Example 7. Additional Assays for Pesticidal Activity

The various polypeptides set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98,99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223,224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279,280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293,294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307,308, and/or 309, or an active variant or fragment of any thereof can betested to act as a pesticide upon a pest in a number of ways. One suchmethod is to perform a feeding assay. In such a feeding assay, oneexposes the pest to a sample containing either compounds to be tested orcontrol samples. Often this is performed by placing the material to betested, or a suitable dilution of such material, onto a material thatthe pest will ingest, such as an artificial diet. The material to betested may be composed of a liquid, solid, or slurry. The material to betested may be placed upon the surface and then allowed to dry.Alternatively, the material to be tested may be mixed with a moltenartificial diet, and then dispensed into the assay chamber. The assaychamber may be, for example, a cup, a dish, or a well of a microtiterplate.

Assays for sucking pests (for example aphids) may involve separating thetest material from the insect by a partition, ideally a portion that canbe pierced by the sucking mouth parts of the sucking insect, to allowingestion of the test material. Often the test material is mixed with afeeding stimulant, such as sucrose, to promote ingestion of the testcompound.

Other types of assays can include microinjection of the test materialinto the mouth, or gut of the pest, as well as development of transgenicplants, followed by test of the ability of the pest to feed upon thetransgenic plant. Plant testing may involve isolation of the plant partsnormally consumed, for example, small cages attached to a leaf, orisolation of entire plants in cages containing insects.

Other methods and approaches to assay pests are known in the art, andcan be found, for example in Robertson and Preisler, eds. (1992)Pesticide bioassays with arthropods, CRC, Boca Raton, Fla.Alternatively, assays are commonly described in the journals ArthropodManagement Tests and Journal of Economic Entomology or by discussionwith members of the Entomological Society of America (ESA). Any one ofSEQ ID NOS: 1-309 can be expressed and employed in an assay as set forthin Examples 3 and 4, herein.

Example 8. Pesticidal Activity Against Coleopteran and Lepidoptera

Protein Expression: Each sequence set forth in Table 3 was expressed inE. coli as described in Example 2. 400 mL of LB was inoculated and grownto an OD600 of 0.6. The culture was induced with 0.25 mM IPTG overnightat 16° C. The cells were spun down and the cell pellet was resuspend in5 mL of buffer. The resuspension was sonicated for 2 min on ice.

Bioassay: Fall army worm (FAW), corn ear worm (CEW), European corn borer(ECB) southwestern corn borer (SWCB) and diamond backed moth (DBM or Px)eggs were purchased from a commercial insectary (Benzon Research Inc.,Carlisle, Pa.). The FAW, CEW, ECB and BCW eggs were incubated to thepoint that eclosion would occur within 12 hrs of the assay setup. SWCBand DBM were introduced to the assay as neonate larvae. Assays werecarried out in 24-well trays containing multispecies lepidopteran diet(Southland Products Inc., Lake Village, Ark.). Samples of the sonicatedlysate were applied to the surface of the diet (diet overlay) andallowed to evaporate and soak into the diet. For CEW, FAW, BCW, ECB andSWCB, a 125 μl of sonicated lysate was added to the diet surface anddried. For DBM, 50 μl of a 1:2 dilution of sonicated lysate was added tothe diet surface. The bioassay plates were sealed with a plate sealingfilm vented with pin holes. The plates were incubated at 26° C. at 65%relative humidity (RH) on a 16:8 day:night cycle in a Percival for 5days. The assays were assessed for level of mortality, growth inhibitionand feeding inhibition.

For the western corn rootworm (WCR) bioassay, the proteinconstruct/lysate was evaluated in an insect bioassay by dispensing 60 μlvolume on the top surface of diet in well/s of 24-well plate (Cellstar,24-well, Greiner Bio One) and allowed to dry. Each well contained 500 μldiet (Marrone et al., 1985). Fifteen to twenty neonate larvae wereintroduced in each well using a fine tip paint brush and the plate wascovered with membrane (Viewseal, Greiner Bio One). The bioassay wasstored at ambient temperature and scored for mortality, and/orgrowth/feeding inhibition at day 4.

For Colorado Potato Beetle (CPB) a cork bore size No. 8 leaf disk wasexcised from potato leaf and was dipped in the protein construct/lysateuntil thoroughly wet and placed on top of filter disk (Millipore, glassfiber filter, 13 mm). 60 μl dH2O was added to each filter disk andplaced in each well of 24-well plate (Cellstar, 24-well, Greiner BioOne). The leaf disk was allowed to dry and five to seven first instarlarvae were introduced in each well using a fine tip paint brush. Theplate was covered with membrane (Viewseal, Greiner Bio One) and smallhole was punctured in each well of the membrane. The construct wasevaluated with four replicates, and scored for mortality and leaf damageon day 3.

Table 3 provides a summary of pesticidal activity against coleopteranand lepidoptera of the various sequences. Table code: “−” indicates noactivity seen; “+” indicates pesticidal activity; “NT” indicates nottested; “S” indicates stunt; “SS” indicates slight stunt; “LF” indicateslow feeding; “M” indicates mortality.

TABLE 3 Summary of Pesticidal Activity against Coleopteran andLepidoptera. WCR APG Seq ID FAW CEW BCW ECB SWCB CPB Px MortalityAPG01199.0 Seq ID 28 M, SS − − NT NT − NT − APG01199.1 Seq ID 29 − − −NT NT − NT − APG01084.0 Seq ID 24 S − S NT NT NT NT − APG01121.0 Seq ID26 − − − NT NT − NT − APG01257.0 Seq ID 34 − − − NT NT − NT − APG00790.0Seq ID 8 − − − NT NT − NT − APG01301.0 Seq ID 37 − − − NT NT − NT −APG01246.1 Seq ID 33 − − − NT NT NT NT − APG00809.0 Seq ID 10 HM, S HM,S HM, S NT NT NT NT APG01150.0 Seq ID 27 SS − − NT NT − NT − APG00788.0Seq ID 7 − − − NT NT − NT − APG00906.0 Seq ID 11 − − − NT NT − NT −APG01401.0 Seq ID 38 − − − NT NT − NT − APG01103.0 Seq ID 25 − − − NT NT− NT − APG00737.0 Seq ID 2 − − − NT NT − NT − APG01269.0 Seq ID 35 SS −− NT NT − NT − APG00989.1 Seq ID 19 S − − NT NT NT NT − APG00808.0 SeqID 9 SS − − NT NT − NT − APG00769.0 Seq ID 6 − − − NT NT − NT −APG00955.1 Seq ID 14 − − − NT NT − NT − APG00749.1 Seq ID 5 − − − NT NT− NT − APG00589.0 Seq ID 1 SS − SS NT NT NT − APG00738.0 Seq ID 3 SS NTNT NT NT − APG00974.0 Seq ID 16 − − − NT NT − NT − APG00974.1 Seq ID 17− − − NT NT NT − APG02531.1 Seq ID 74 − − − NT NT NT NT − APG03114.1 SeqID 92 − − − NT NT NT NT − APG02921.1 Seq ID 88 − − − NT NT − NT −APG09455.1 Seq ID 203 S − − NT NT NT NT − APG07444.0 Seq ID 167 − − − NTNT − NT − APG06997.0 Seq ID 157 SS NT NT NT NT − APG07224.1 Seq ID 166 −− − NT NT NT NT − APG09376.0 Seq ID 200 − − − NT NT − NT − APG09376.1Seq ID 201 − − − NT NT − NT − APG07639.0 Seq ID 171 SS − − NT NT NT −APG01451.0 Seq ID 41 − − − NT NT − NT − APG05500.1 Seq ID 131 M, SS − −NT NT NT NT − APG02067.2 Seq ID 61 M, S − − NT NT + NT − APG01992.1 SeqID 57 M, S − − NT NT + NT − APG06324.1 Seq ID 140 − − − NT NT − NT −APG05706.1 Seq ID 135 − − − NT NT + NT − APG01700.1 Seq ID 51 S − S NTNT − NT − APG07220.0 Seq ID 164 − − − NT NT NT NT − APG09659.1 Seq ID207 S − − NT NT − NT − APG08780.1 Seq ID 193 − − − NT NT − NT −APG00945.0 Seq ID 12 SS − NT NT − NT − APG09717.0 Seq ID 208 − − − NT NTNT NT − APG08509.1 Seq ID 186 M, SS S SS NT NT + NT + APG05553.0 Seq ID132 SS NT NT NT NT − APG01068.1 Seq ID 22 − − − NT NT − NT − APG07682.0Seq ID 174 − − − NT NT + NT − APG07682.1 Seq ID 175 − − − NT NT − NT −APG06465.0 Seq ID 146 S − − NT NT − NT − APG04793.1 Seq ID 125 SS − SSNT NT − NT − APG06989.2 Seq ID 155 − − − NT NT − NT − APG07470.0 Seq ID170 S − SS NT NT − NT − APG03619.1 Seq ID 100 − − − NT NT − NT −APG02923.0 Seq ID 89 − − − NT NT + NT − APG04686.1 Seq ID 121 SS − − NTNT − NT − APG01536.1 Seq ID 48 − − − NT NT NT NT − APG02555.0 Seq ID 77− − − NT NT − NT − APG02555.1 Seq ID 78 − − − NT NT − NT − APG01463.0Seq ID 42 S − − NT NT + NT − APG06338.1 Seq ID 142 SS − − NT NT − NT −APG04152.1 Seq ID 117 − − − NT NT NT NT − APG03440.0 Seq ID 95 S SS NTNT NT NT − APG02225.0 Seq ID 64 − − − NT NT NT NT − APG01508.1 Seq ID 46− − − NT NT NT NT − APG02518.1 Seq ID 72 − − − NT NT − NT − APG09642.1Seq ID 205 − − − NT NT NT NT − APG02740.1 Seq ID 85 SS − − NT NT − NT −APG08628.1 Seq ID 191 S − − NT NT − NT − APG03238.1 Seq ID 94 − − − NTNT NT NT − APG03831.1 Seq ID 109 M, S − S NT NT NT NT − APG05213.0 SeqID 127 M, S − S NT NT NT NT − APG02552.1 Seq ID 76 − − − NT NT NT NT −APG03484.2 Seq ID 98 SS − − NT NT − NT − APG01242.1 Seq ID 31 − − − NTNT NT NT − APG04099.0 Seq ID 114 − − − NT NT NT NT − APG04099.1 Seq ID115 − − − NT NT − NT − APG08151.1 Seq ID 184 S SS − NT NT NT NT −APG09735.1 Seq ID 212 M, S − − NT NT − NT − APG06501.0 Seq ID 147 M, S −− NT NT + NT − APG06501.1 Seq ID 148 − − − NT NT − NT − APG08794.0 SeqID 194 − − − NT NT + NT − APG08794.1 Seq ID 195 − − − NT NT − NT −APG01078.0 Seq ID 23 − − − NT NT − NT − APG05634.0 Seq ID 133 SS − − NTNT − NT − APG03715.1 Seq ID 104 M, S − − NT NT − NT − APG07002.1 Seq ID159 − − − NT NT − NT − APG08607.2 Seq ID 188 SS SS NT NT − NT −APG04925.0 Seq ID 126 − − − NT NT NT NT − APG04721.0 Seq ID 122 SS − SSNT NT NT NT − APG02400.1 Seq ID 70 − − − NT NT NT NT − APG09096.0 Seq ID198 M, S − − NT NT − NT − APG09096.1 Seq ID 199 − − − NT NT NT NT −APG02280.0 Seq ID 65 − − − NT NT − NT − APG01507.1 Seq ID 44 − − − NT NT− NT − APG03686.1 Seq ID 102 SS − SS NT NT − NT − APG04483.0 Seq ID 119S − − NT NT − NT − APG03747.1 Seq ID 106 − − − NT NT − NT − APG01882.1Seq ID 53 − − − NT NT − NT − APG06281.0 Seq ID 138 − − − NT NT − NT −APG02643.1 Seq ID 83 − − − NT NT − NT − APG06894.1 Seq ID 153 S − − NTNT − NT − APG02038.0 Seq ID 58 − − − NT NT − NT − APG03760.0 Seq ID 107HM, S HM, S HM, S NT NT + NT + APG06676.0 Seq ID 151 − − − NT NT NT NT −APG09726.1 Seq ID 210 − − − NT NT NT NT − APG05372.1 Seq ID 129 − − − NTNT NT NT − APG07114.0 Seq ID 162 − − − NT NT + NT − APG07114.1 Seq ID163 − − − NT NT − NT − APG04067.0 Seq ID 112 M, S − − NT NT + NT −APG04067.1 Seq ID 113 − − − NT NT NT NT − APG06381.2 Seq ID 144 − − − NTNT − NT − APG08029.1 Seq ID 181 − − − NT NT − NT − APG04778.0 Seq ID 123S − − NT NT + NT − APG02387.2 Seq ID 67 S − S NT NT − NT − APG02557.0Seq ID 79 S − S NT NT − NT − APG02557.1 Seq ID 80 − − − NT NT NT NT −APG06589.1 Seq ID 150 SS − − NT NT − NT − APG07780.0 Seq ID 178 − − − NTNT − NT − APG02633.0 Seq ID 81 SS − − NT NT − NT − APG04450.0 Seq ID 118M, SS − − NT NT NT NT − APG07676.1 Seq ID 173 S SS SS NT NT − NT −APG08138.0 Seq ID 182 S − − NT NT − NT − APG06001.1 Seq ID 137 SS SS −NT NT NT NT − APG01420.0 Seq ID 39 − − − NT NT NT NT − APG01420.1 Seq ID40 SS − − NT NT − NT − APG03867.1 Seq ID 111 − − − NT NT NT NT −APG09055.0 Seq ID 196 M, S − − NT NT NT NT − APG09055.1 Seq ID 197 − − −NT NT − NT − APG01273.0 Seq ID 36 − − − NT NT − NT − APG07738.1 Seq ID177 − − − NT NT NT NT − APG03079.0 Seq ID 90 NT NT NT NT NT − NT −APG05660.1 Seq ID 245 S − S NT NT NT NT − APG03217.1 Seq ID 230 M, S − SNT NT NT NT − APG08085.1 Seq ID 257 M, S − S NT NT NT NT − APG08973.1Seq ID 265 − − − NT NT NT NT − APG03185.1 Seq ID 228 S S S NT NT − NT −APG08241.0 Seq ID 260 S − − NT NT NT NT + APG05969.0 Seq ID 246 HM, S −− NT NT NT NT − APG04226.1 Seq ID 238 M, S − − NT NT NT NT − APG01705.1Seq ID 217 M, S − − NT NT NT NT − APG07049.1 Seq ID 251 M, S − − NT NTNT NT − APG08990.1 Seq ID 267 M, S − S NT NT NT NT − APG03368.1 Seq ID232 M, S − − NT NT NT NT − APG02768.0 Seq ID 225 SS − S NT NT NT NT −APG09842.0 Seq ID 269 M, S − S NT NT NT NT − APG01989.0 Seq ID 218 M, S− S NT NT NT NT − APG02245.0 Seq ID 219 M, S − S NT NT NT NT −APG08718.0 Seq ID 263 M, S − S NT NT NT NT − APG02429.0 Seq ID 222 M, S− S NT NT NT NT − APG00743.1 Seq ID 215 M, S − S NT NT NT NT −APG04643.2 Seq ID 242 M, S − S NT NT NT NT − APG03040.0 Seq ID 226 M, S− S NT NT NT NT − APG09256.0 Seq ID 268 M, S − − NT NT + NT − APG02674.0Seq ID 223 S − − NT NT NT NT − APG02674.1 Seq ID 224 M, S − − NT NT − NT− APG03662.0 Seq ID 233 S SS SS NT NT − NT − APG08043.1 Seq ID 255 M, SS − NT NT − NT − APG08411.0 Seq ID 261 SS − S NT NT NT NT − APG08411.1Seq ID 262 S − − NT NT − NT − APG07574.0 Seq ID 252 − − − NT NT − NT −APG07574.1 Seq ID 253 S − − NT NT − NT − APG04485.1 Seq ID 240 S − − NTNT − NT − APG08225.1 Seq ID 259 S − S NT NT − NT − APG02960.2 Seq ID 272S − − NT NT − NT − APG01172.2 Seq ID 303 SS − − NT NT NT NT − APG05711.0Seq ID 307 HM, S − − NT NT − NT − APG05711.1 Seq ID 308 M, S − S NT NT +NT − APG01288.0 Seq ID 306 S − − NT NT + NT − APG08085.0 Seq ID 256 − −NT NT NT NT NT − APG04226.0 Seq ID 237 − − NT NT NT NT NT − APG02248.0Seq ID 284 SS − NT NT NT NT NT − APG09857.0 Seq ID 300 SS − NT NT NT NTNT − APG03574.1 Seq ID 290 SS SS NT NT NT NT NT − APG05678.0 Seq ID 292SS S NT NT NT NT NT − APG03148.0 Seq ID 288 SS − NT NT NT NT NT −APG01577.0 Seq ID 281 − − NT NT NT NT NT − APG07445.1 Seq ID 169 − − −NT NT − NT − APG01022.0 Seq ID 20 − − − NT NT − NT − APG07020.1 Seq ID161 SS − SS NT NT − NT − APG06502.0 Seq ID 4 S − − NT NT − NT −APG01705.0 Seq ID 216 SS − − NT NT − NT −

Example 9. Pesticidal Activity Against Hemipteran

Protein Expression: Each of the sequences set forth in Table 4 wasexpressed in E. coli as described in Example 2. 400 mL of LB wasinoculated and grown to an OD600 of 0.6. The culture was induced with0.25 mM IPTG overnight at 16° C. The cells were spun down and the cellpellet was re-suspend in 5 mL of buffer. The resuspension was sonicatedfor 2 min on ice.

Second instar southern green stinkbug (SGSB) were obtained from acommercial insectary (Benzon Research Inc., Carlisle, Pa.). A 50% v/vratio of sonicated lysate sample to 20% sucrose was employed in thebioassay. Stretched parafilm was used as a feeding membrane to exposethe SGSB to the diet/sample mixture. The plates were incubated at 25°C.: 21° C., 16:8 day:night cycle at 65% RH for 5 days.

Mortality was scored for each sample. The results are set forth in Table4. A dashed line indicates no mortality was detected. The proteinslisted in Table 4 showed from about 10% to about 100% mortality 25%mortality or 50% mortality (as indicated) against southern greenstinkbug (1 stinkbug out of 4 died). The negative controls (empty vectorexpressed binding domain and buffer only) both showed no mortality (0stinkbugs out of 4).

TABLE 4 Summary of Pesticidal Activity against Hemipteran APG Seq IDTested against SGSB APG08509.1 Seq ID 186 100%  APG03185.1 Seq ID 228100%  APG02225.0 Seq ID 64 90% APG03831.1 Seq ID 109 90% APG03760.0 SeqID 107 90% APG07574.1 Seq ID 253 90% APG08151.1 Seq ID 184 80%APG03662.0 Seq ID 233 80% APG03238.1 Seq ID 94 70% APG03079.0 Seq ID 9070% APG05213.0 Seq ID 127 60% APG02552.1 Seq ID 76 60% APG04643.2 Seq ID242 60% APG01084.0 Seq ID 24 50% APG01150.0 Seq ID 27 50% APG00788.0 SeqID 7 50% APG00906.0 Seq ID 11 50% APG03114.1 Seq ID 92 50% APG07639.0Seq ID 171 50% APG07470.0 Seq ID 170 50% APG08607.2 Seq ID 188 50%APG09096.0 Seq ID 198 50% APG09842.0 Seq ID 269 50% APG02674.0 Seq ID223 50% APG01172.2 Seq ID 303 50% APG00738.0 Seq ID 3 40% APG07682.0 SeqID 174 40% APG06338.1 Seq ID 142 40% APG02387.2 Seq ID 67 40% APG08085.1Seq ID 257 40% APG08241.0 Seq ID 260 40% APG07049.1 Seq ID 251 40%APG08990.1 Seq ID 267 40% APG02768.0 Seq ID 225 40% APG08411.0 Seq ID261 40% APG07574.0 Seq ID 252 40% ApG05711.1 Seq ID 308 40% APG02923.0Seq ID 89 33% APG02067.2 Seq ID 61 30% APG01992.1 Seq ID 57 30%APG05706.1 Seq ID 135 30% APG01700.1 Seq ID 51 30% APG04099.1 Seq ID 11530% APG06501.0 Seq ID 147 30% APG05634.0 Seq ID 133 30% APG04721.0 SeqID 122 30% APG09096.1 Seq ID 199 30% APG07114.0 Seq ID 162 30%APG05969.0 Seq ID 246 30% APG03368.1 Seq ID 232 30% APG02245.0 Seq ID219 30% APG00743.1 Seq ID 215 30% APG03040.0 Seq ID 226 30% APG02674.1Seq ID 224 30% APG04485.1 Seq ID 240 30% APG05711.0 Seq ID 307 30%APG01288.0 Seq ID 306 30% APG01121.0 Seq ID 26 25% APG00790.0 Seq ID 825% APG01301.0 Seq ID 37 25% APG01103.0 Seq ID 25 25% APG00769.0 Seq ID6 25% APG02531.1 Seq ID 74 25% APG07444.0 Seq ID 167 25% APG06324.1 SeqID 140 25% APG09717.0 Seq ID 208 25% APG01068.1 Seq ID 22 25% APG01536.1Seq ID 48 25% APG02555.0 Seq ID 77 25% APG09642.1 Seq ID 205 25%APG08628.1 Seq ID 191 25% APG08794.1 Seq ID 195 25% APG02400.1 Seq ID 7025% APG01882.1 Seq ID 53 25% APG06281.0 Seq ID 138 25% APG09726.1 Seq ID210 25% APG04067.1 Seq ID 113 25% APG06381.2 Seq ID 144 25% APG07780.0Seq ID 178 25% APG02633.0 Seq ID 81 25% APG04450.0 Seq ID 118 25%APG03867.1 Seq ID 111 25% APG09055.1 Seq ID 197 25% APG01273.0 Seq ID 3625% APG00955.1 Seq ID 14 20% APG07220.0 Seq ID 164 20% APG05553.0 Seq ID132 20% APG04686.1 Seq ID 121 20% APG03484.2 Seq ID 98 20% APG02280.0Seq ID 65 20% APG02557.0 Seq ID 79 20% APG01420.0 Seq ID 39 50%APG09055.0 Seq ID 196 20% APG03217.1 Seq ID 230 20% APG01989.0 Seq ID218 20% APG09256.0 Seq ID 268 20% APG08043.1 Seq ID 255 20% APG08411.1Seq ID 262 20% APG02960.2 Seq ID 272 20% APG07682.1 Seq ID 175 10%APG04099.0 Seq ID 114 10% APG09735.1 Seq ID 212 10% APG08794.0 Seq ID194 10% APG04925.0 Seq ID 126 10% APG04067.0 Seq ID 112 10% APG05660.1Seq ID 245 10% APG08973.1 Seq ID 265 10% APG08718.0 Seq ID 263 10%APG02429.0 Seq ID 222 10% APG08225.1 Seq ID 259 10% APG08085.0 Seq ID256 60% APG04226.0 Seq ID 237 70% APG02248.0 Seq ID 284 50% APG09857.0Seq ID 300 40% APG03574.1 Seq ID 290 40% APG05678.0 Seq ID 292 20%APG03148.0 Seq ID 288 40% APG01577.0 Seq ID 281 10% APG07445.1 Seq ID169 10% APG06502.0 Seq ID 4 20% APG00589.0 Seq ID 1 60%

SEQ ID NOs. 2, 5, 9, 10, 12, 16, 17, 19, 20, 23, 28, 29, 31, 33, 34, 35,38, 40, 41, 42, 44, 46, 58, 72, 78, 80, 83, 85, 88, 95, 100, 102, 104,106, 117, 119, 123, 125, 129, 131, 137, 140, 146, 148, 150, 151, 153,155, 157, 159, 161, 163, 166, 173, 177, 181, 182, 193, 200, 201, 203,207, 216, 217 and 238 were tested and did not have activity in thisexperiment.

Example 10. Pesticidal Activity Against Soybean Aphid

Protein Expression: Each sequence set forth in SEQ ID NOS: 1-309 (or anactive variant or fragment thereof) is expressed in E. coli as describedin Example 2. 400 mL of LB is inoculated and grown to an OD600 of 0.6.The culture is induced with 0.25 mM IPTG overnight at 16° C. The cellsare spun down and the cell pellet is resuspend in 5 mL of buffer. Theresuspension is sonicated for 2 min on ice.

Soybean aphids (SBA) are obtained from Michigan State University. Sixadult aphids are added to each well of a 24 well plate. Purifiedproteins are provided in liquid artificial diet at a rate of 25% (50 ulprotein, 150 ul artificial diet), and are sealed with an artificialmembrane through which the aphids are able to feed. The plates are heldin an incubator at 26° C., 60% RH, 16:8 day:night cycle for 5 days.

Mortality, feeding, and reproduction are scored for each sample on days3, 4, and 5 post-treatment. Mortality is calculated as percent dead ofthe original 6 adult aphids. Feed and reproduction activities areassigned a score on a 0-3 point scale with 0 being no feeding orreproduction and 3 being high feeding or reproduction. Feeding ismeasured as the amount of honeydew (liquid excretions produced byaphids) that collected in each well. Reproduction is the number of liveimmature aphids that are observed in each well. Mortality, feeding, andreproduction data are assessed using a combined scoring system toestablish cutoff levels for activity. The combined score is calculatedas: Combined Score=(Feeding+Reproduction+Mortality Score)/3, whereMortality Score=3−(3×% Mortality/100).

TABLE 5 Cut-offs used to rate individual wells as active or not for eachobservational day post-introduction of aphids to assay wells. A well isdeemed active if measure ≤ cut-off. Cut-off used to designate activityActivity Measure Day 3 Day 4 Day 5 Combined Score 2 1.5 1

Example 11. Pesticidal Activity Against Soybean Aphid

Protein Expression: Each sequence set forth in Table 7 (or an activevariant or fragment thereof) was expressed in E. coli as described inExample 2. 400 mL of LB was inoculated and grown to an OD600 of 0.6. Theculture was induced with 0.25 mM IPTG overnight at 16° C. The cells werespun down and the cell pellet was resuspend in 5 mL of buffer. Theresuspension was sonicated for 2 min on ice.

Soybean aphids (SBA) were obtained from Michigan State University. Sixadult aphids were added to each well of a 24 well plate. Purifiedproteins were provided in liquid artificial diet at a rate of 25% (50 ulprotein, 150 ul artificial diet), and were sealed with an artificialmembrane through which the aphids are able to feed. The plates were heldin an incubator at 26° C., 60% RH, 16:8 day:night cycle for 5 days.

Mortality, feeding, and reproduction were scored for each sample on days3, 4, and 5 post-treatment. Mortality was calculated as percent dead ofthe original 6 adult aphids. Feed and reproduction activities wereassigned a score on a 0-3 point scale with 0 being no feeding orreproduction and 3 being high feeding or reproduction. Feeding wasmeasured as the amount of honeydew (liquid excretions produced byaphids) that collected in each well. Reproduction was the number of liveimmature aphids that were observed in each well. Mortality, feeding, andreproduction data were assessed using a combined scoring system toestablish cutoff levels for activity. The combined score was calculatedas:

Combined Score=(Feeding+Reproduction+Mortality Score)/3, where MortalityScore=3−(3×% Mortality/100). The results are set forth in Table 7. “+”indicates pesticidal activity.

TABLE 6 Cut-offs used to rate individual wells as active or not for eachobservational day post-introduction of aphids to assay wells. A well isdeemed active if measure ≤ cut-off. Cut-off used to designate activityActivity Measure Day 3 Day 4 Day 5 Combined Score 2 1.5 1

TABLE 7 Summary of Pesticidal Activity against soybean aphid APG Seq IDTested against SBA APG08509.1 Seq ID 186 + APG08628.1 Seq ID 191 +APG03185.1 Seq ID 228 + APG07574.1 Seq ID 253 +

SEQ ID NOs: 1, 3, 4, 5, 14, 19, 20, 23, 24, 29, 31, 36, 39, 40, 41, 42,44, 46, 48, 51, 53, 57, 61, 64, 65, 67, 70, 72, 74, 76, 77, 78, 79, 80,83, 88, 89, 90, 94, 95, 98, 100, 104, 107, 109, 112, 113, 114, 115, 118,119, 121, 122, 123, 126, 127, 129, 132, 133, 135, 137, 138, 140, 142,144, 146, 147, 148, 150, 151, 153, 155, 157, 159, 161, 162, 163, 164,166, 169, 170, 174, 175, 177, 181, 182, 184, 188, 193, 194, 195, 196,198, 199, 200, 201, 203, 205, 207, 212, 215, 216, 217, 218, 219, 222,223, 224, 225, 226, 230, 232, 233, 237, 238, 240, 242, 245, 246, 251,252, 255, 256, 257, 259, 260, 261, 262, 263, 265, 267, 268, 269, 272,281, 284, 288, 290, 292, 300, 306, 307 and 308 were tested and did nothave activity in this experiment.

All publications and patent applications mentioned in the specificationare indicative of the level of skill of those skilled in the art towhich this invention pertains. All publications and patent applicationsare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the appended claims.

That which is claimed is:
 1. A recombinant polypeptide having pesticidalactivity, comprising: (a) a polypeptide comprising an amino acidsequence having at least 90% percent sequence identity to an amino acidsequence selected from the group consisting of sequences set forth inSEQ ID NOs: 107, 228, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51,52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104,105, 106, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119,120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133,134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147,148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161,162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175,176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189,190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203,204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217,218, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 229, 230, 231,232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245,246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259,260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273,274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287,288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301,302, 303, 304, 305, 306, 307, 308, or 309; or, (b) a polypeptidecomprising the amino acid sequence set forth in SEQ ID NOs: 107, 228, 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39,40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 108, 109,110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123,124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137,138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151,152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165,166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179,180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193,194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207,208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220,221, 222, 223, 224, 225, 226, 227, 229, 230, 231, 232, 233, 234, 235,236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249,250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263,264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291,292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305,306, 307, 308, or
 309. 2. The polypeptide of claim 1, further comprisinga heterologous amino acid sequence.
 3. A composition comprising thepolypeptide of claim
 1. 4. A recombinant nucleic acid molecule encodingan amino acid sequence comprising: (a) at least 90% percent sequenceidentity to an amino acid sequence selected from the group consisting ofsequences set forth in SEQ ID NOs: 107, 228, 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254,255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268,269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282,283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, or 309; or(b) the amino acid sequence set forth in SEQ ID NOs: 107, 228, 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58,59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76,77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 108, 109, 110,111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124,125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138,139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152,153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166,167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180,181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194,195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208,209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221,222, 223, 224, 225, 226, 227, 229, 230, 231, 232, 233, 234, 235, 236,237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250,251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264,265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278,279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292,293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306,307, 308, or 309; wherein said recombinant nucleic acid molecule is nota naturally occurring sequence encoding said polypeptide.
 5. Therecombinant nucleic acid molecule of claim 4, wherein said recombinantnucleic acid molecule is a synthetic sequence designed for expression ina plant.
 6. The recombinant nucleic acid molecule of claim 4, whereinsaid recombinant nucleic acid molecule is operably linked to a promotercapable of directing expression in a plant cell.
 7. The recombinantnucleic acid molecule of claim 4, wherein said recombinant nucleic acidmolecule is operably linked to a promoter capable of directingexpression in a bacteria.
 8. A host cell comprising the recombinantnucleic acid molecule of claim
 4. 9. The host cell of claim 8, whereinsaid host cell is a bacterial host cell.
 10. A DNA construct comprisinga promoter that drives expression in a plant cell operably linked to arecombinant nucleic acid molecule comprising a nucleotide sequence thatencodes a polypeptide comprising an amino acid sequence having at least90% percent sequence identity to an amino acid sequence selected fromthe group consisting of sequences set forth in SEQ ID NOs: 107, 228, 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39,40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 108, 109,110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123,124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137,138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151,152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165,166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179,180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193,194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207,208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220,221, 222, 223, 224, 225, 226, 227, 229, 230, 231, 232, 233, 234, 235,236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249,250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263,264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291,292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305,306, 307, 308, or
 309. 11. The DNA construct of claim 10, wherein saidnucleotide sequence is a synthetic DNA sequence designed for expressionin a plant.
 12. A vector comprising the DNA construct of claim
 10. 13. Ahost cell comprising the vector of claim
 12. 14. A compositioncomprising the host cell of claim
 13. 15. The composition of claim 14,wherein said composition is selected from the group consisting of apowder, dust, pellet, granule, spray, emulsion, colloid, and solution.16. The composition of claim 15, wherein said composition comprises fromabout 1% to about 99% by weight of said polypeptide.
 17. A method forcontrolling a pest population comprising contacting said pest populationwith a pesticidal-effective amount of the composition of claim
 3. 18. Amethod for producing a polypeptide with pesticidal activity comprisingculturing the host cell of claim 13 under conditions in which thenucleic acid molecule encoding the polypeptide is expressed.
 19. A planthaving stably incorporated into its genome a DNA construct comprising anucleotide sequence that encodes a protein having pesticidal activity,wherein said nucleotide sequence comprises: (a) a nucleotide sequencethat encodes a polypeptide comprising the amino acid sequence of any oneof SEQ ID NOs: 107, 228, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50,51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103,104, 105, 106, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118,119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132,133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146,147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160,161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174,175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188,189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202,203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216,217, 218, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 229, 230,231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244,245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258,259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272,273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286,287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300,301, 302, 303, 304, 305, 306, 307, 308, or 309; or (b) a nucleotidesequence that encodes a polypeptide comprising an amino acid sequencehaving at least 90% percent sequence identity to an amino acid sequenceselected from the group consisting of sequences set forth in SEQ ID NOs:107, 228, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36,37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72,73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106,108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121,122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135,136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149,150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163,164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177,178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191,192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205,206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 218,219, 220, 221, 222, 223, 224, 225, 226, 227, 229, 230, 231, 232, 233,234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247,248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261,262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275,276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289,290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303,304, 305, 306, 307, 308, or
 309. 20. A transgenic seed of the plant ofclaim
 19. 21. A method for protecting a plant from an insect pest,comprising expressing in a plant or cell thereof a nucleotide sequencethat encodes a pesticidal polypeptide, wherein said nucleotide sequencecomprising: (a) a nucleotide sequence that encodes a polypeptidecomprising the amino acid sequence of any one of SEQ ID NOs: 107, 228,1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38,39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56,57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 108, 109,110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123,124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137,138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151,152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165,166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179,180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193,194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207,208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220,221, 222, 223, 224, 225, 226, 227, 229, 230, 231, 232, 233, 234, 235,236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249,250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263,264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291,292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305,306, 307, 308, or 309; or (b) a nucleotide sequence that encodes apolypeptide comprising an amino acid sequence having at least 90%percent sequence identity to an amino acid sequence selected from thegroup consisting of sequences set forth in SEQ ID NOs: 107, 228, 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58,59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76,77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 108, 109, 110,111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124,125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138,139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152,153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166,167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180,181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194,195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208,209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 218, 219, 220, 221,222, 223, 224, 225, 226, 227, 229, 230, 231, 232, 233, 234, 235, 236,237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250,251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264,265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278,279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292,293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306,307, 308, or
 309. 22. The method of claim 21, wherein said plantproduces a pesticidal polypeptide having pesticidal activity against atleast one of a lepidopteran pest, a coleopteran pest, or a hemipteranpest.
 23. A method for increasing yield in a plant comprising growing ina field a plant or seed thereof having stably incorporated into itsgenome a DNA construct comprising a promoter that drives expression in aplant operably linked to a nucleotide sequence that encodes a pesticidalpolypeptide, wherein said nucleotide sequence comprises: (a) anucleotide sequence that encodes a polypeptide comprising the amino acidsequence of any one of SEQ ID NOs: 107, 228, 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128,129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156,157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170,171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184,185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 218, 219, 220, 221, 222, 223, 224, 225,226, 227, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254,255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268,269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282,283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, or 309; or anucleotide sequence that encodes a polypeptide comprising an amino acidsequence having at least 90% percent sequence identity to an amino acidsequence selected from the group consisting of sequences set forth inSEQ ID NOs: 107, 228, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51,52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104,105, 106, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119,120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133,134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147,148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161,162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175,176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189,190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203,204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217,218, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 229, 230, 231,232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245,246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259,260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273,274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287,288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301,302, 303, 304, 305, 306, 307, 308, or 309.